TY  - JOUR
AB  - Equine recurrent uveitis (ERU) is a spontaneous, remitting-relapsing autoimmune disease driven by the adaptive immune system. Although T cells are described as the main effector cells in pathogenesis, granulocytes have also emerged as possible disease mediators. To explore the role of these innate immune cells, we investigated the whole cell proteome of granulocytes from equine recurrent uveitis cases and healthy controls. Among the 2362 proteins identified by mass spectrometry, we found 96 proteins with significantly changed abundance between groups (p < 0.05, fold change >1.2), representing 4.1% of total granulocyte proteome. Within these differential identifications, calgranulin B, a protein associated with pathogenesis in other autoimmune diseases, showed highest abundance in equine recurrent uveitis (18 fold). For a better interpretation of the results from our hypothesis-generating approach, we added a threshold for biological significance (ratio ERU/controls >2: 36 proteins) to the proteins with increased abundance in equine recurrent uveitis and analyzed their allocation to the subsets within the Immune System superpathway. The 36 differentially abundant proteins predominantly associated to RAF/MAP kinase cascade, MHC-I-mediated antigen presentation and neutrophil degranulation, suggesting a latently activated phenotype of these innate immune cells in disease. Raw data are available via ProteomeXchange with identifier PXD013648. Significance: Our study provides new insights into the protein repertoire of primary equine granulocytes and identifies protein abundance changes associated to equine recurrent uveitis (ERU), an organ specific, spontaneously occurring autoimmune disease. We show that granulocyte proteins with increased abundance in ERU strongly associate to RAF/MAP kinase signaling, MHC-I antigen presentation and neutrophil degranulation, pointing to a more activated state of these cells in ERU cases. Since cells were obtained in quiescent stage of disease, latent activation of granulocytes underlines the role of these innate immune cells in ERU. These findings are highly relevant for veterinary medicine, further establishing the importance of granulocytes in this T cell-driven autoimmune disease. Moreover, they have translational quality for autoimmune uveitis in man, due to strong similarity in disease occurrence, progression and pathogenesis.
AU  - Weigand, M.*
AU  - Hauck, S.M.
AU  - Deeg, C.A.*
AU  - Degroote, R.L.*
C1  - 60263
C2  - 49303
CY  - Radarweg 29, 1043 Nx Amsterdam, Netherlands
TI  - Deviant proteome profile of equine granulocytes associates to latent activation status in organ specific autoimmune disease.
JO  - J. Proteomics
VL  - 230
PB  - Elsevier
PY  - 2021
SN  - 1874-3919
ER  - 

TY  - JOUR
AB  - Nephrotic syndrome is characterized by urinary excretion of plasma proteases or proteasuria. There is a lack of data on the quantity, activity status and identity of these aberrantly filtered proteases. We established a fluorescence-based substrate assay to quantify protease activity in urine samples from healthy and nephrotic humans and mice. Protease class activity was determined after addition of specific inhibitors. Individual proteases were identified by tandem mass spectrometry (MS/MS). In spot urine samples from 10 patients with acute nephrotic syndrome of various etiology, urinary protease activity was significantly increased compared to that of healthy persons (753 ± 178 vs. 244 ± 65 relative units, p < 0.05). The corresponding proteases were highly sensitive to inhibition by the serine protease inhibitors AEBSF (reduction by 85 ± 6% and 72 ± 8%, respectively) and aprotinin (83 ± 9% vs. 25 ± 6%, p < 0.05). MS/MS of all urinary proteins or after AEBSF purification showed that most of them were active serine proteases from the coagulation and complement cascade. These findings were recapitulated in mice, pointing to a similar pathophysiology. In conclusion, nephrotic syndrome leads to increased urinary excretion of active plasma proteases which can be termed proteasuria. Serine proteases account for the vast majority of urinary protease activity in health and nephrotic syndrome. Significance statement: In this study, we found that nephrotic urine samples of humans and mice have a significantly increased protease activity compared to healthy urine samples, using a universal pentapeptide substrate library. This was driven by increased excretion of aprotinin-sensitive serine proteases. With tandem mass spectrometry, we provide a comprehensive and systematic overview of all urinary proteases or the “urine proteasome”. We identified renally expressed proteases in health and addition of proteases from the coagulation and complement cascade in the nephrotic state. These results set the basis to study the role of urinary proteases at both health and nephrotic syndrome to find diagnostic markers of renal disease as well as possible therapeutic targets.
AU  - Wörn, M.*
AU  - Bohnert, B.N.
AU  - Alenazi, F.*
AU  - Boldt, K.*
AU  - Klose, F.*
AU  - Junger, K.*
AU  - Ueffing, M.*
AU  - Birkenfeld, A.L.
AU  - Kalbacher, H.*
AU  - Artunc, F.
C1  - 60145
C2  - 49273
CY  - Radarweg 29, 1043 Nx Amsterdam, Netherlands
TI  - Proteasuria in nephrotic syndrome–quantification and proteomic profiling.
JO  - J. Proteomics
VL  - 230
PB  - Elsevier
PY  - 2021
SN  - 1874-3919
ER  - 

TY  - JOUR
AB  - In life sciences, antibodies are among the most commonly used tools for identifying, tracking, quantifying and isolating molecules, mainly proteins. However, it has recently become clear that antibodies often fall short with respect to specificity and selectivity and in many cases target proteins are not even known. When commercial availability of antibodies is scarce, e.g. for targeting proteins from farm animals, researchers face additional challenges: they often have to rely on cross-reactive antibodies, which are poorly characterized for their exact target, their actual cross-reactivity and the desired application.In this study, we aimed at identifying the true target of mouse monoclonal antibody 8F2, which was generated against chicken PBMC and used for decades in research, while it's actual target molecule remained unknown. We used 8F2 antibody for immunoprecipitation in chicken PBMC and subsequently identified its true target as CD11d, which was never described in chicken lymphocytes before, by quantitative LC-MSMS. The most abundant interactor of CD11d was identified as integrin beta 2.The existence of this alpha integrin was therefore clearly proven on protein level and provides a first basis to further assess the role of CD11d in chickens in future studies.Data are available via ProteomeXchange with identifier PXD017248.Significance: Our studies determined CD11d as the true target of a previously uncharacterized mouse monoclonal antibody 8F2, generated against chicken peripheral blood derived mononuclear cells (PBMC). This is therefore now first member of alpha integrins in chickens, that existence was now clearly identified on protein level. The additional identification of CD11d interactors provides information on integrin-dependent regulation of signaling networks, allowing further functional studies.
AU  - Deeg, C.A.*
AU  - Degroote, R.L.*
AU  - Giese, I.M.*
AU  - Hirmer, S.*
AU  - Amann, B.*
AU  - Weigand, M.*
AU  - Wiedemann, C.*
AU  - Hauck, S.M.
C1  - 59466
C2  - 48863
CY  - Radarweg 29, 1043 Nx Amsterdam, Netherlands
TI  - CD11d is a novel antigen on chicken leukocytes.
JO  - J. Proteomics
VL  - 225
PB  - Elsevier
PY  - 2020
SN  - 1874-3919
ER  - 

TY  - JOUR
AB  - INSC94Y transgenic pigs develop a stable diabetic phenotype early after birth and therefore allow studying the influence of hyperglycemia on primary immune cells in an early stage of diabetes mellitus in vivo. Since immune response is altered in diabetes mellitus, with deviant neutrophil function discussed as one of the possible causes in humans and mouse models, we investigated these immune cells in INSC94Y transgenic pigs and wild type controls at protein level. A total of 2371 proteins were quantified by label-free LC-MS/MS. Subsequent differential proteome analysis of transgenic animals and controls revealed clear differences in protein abundances, indicating a deviant behavior of granulocytes in the diabetic state. Interestingly, abundance of myosin regulatory light chain 9 (MLC-2C) was increased 5-fold in cells of diabetic pigs. MLC-2C directly affects cell contractility by regulating myosin ATPase activity, can act as transcription factor and was also associated with inflammation. It might contribute to impaired neutrophil cell adhesion, migration and phagocytosis.Our study provides novel insights into proteome changes in neutrophils from a large animal model for permanent neonatal diabetes mellitus and points to dysregulation of neutrophil function even in an early stage of this disease.Data are available via ProteomeXchange with identifier PXD017274.Significance: Our studies provide novel basic information about the neutrophil proteome of pigs and contribute to a better understanding of molecular mechanisms involved in altered immune cell function in an early stage diabetes. We demonstrate proteins that are dysregulated in neutrophils from a transgenic diabetic pig and have not been described in this context so far. The data presented here are highly relevant for veterinary medicine and have translational quality for diabetes in humans.
AU  - Weigand, M.*
AU  - Degroote, R.L.*
AU  - Amann, B.*
AU  - Renner, S.*
AU  - Wolf, E.*
AU  - Hauck, S.M.
AU  - Deeg, C.A.*
C1  - 59290
C2  - 48773
CY  - Radarweg 29, 1043 Nx Amsterdam, Netherlands
TI  - Proteome profile of neutrophils from a transgenic diabetic pig model shows distinct changes.
JO  - J. Proteomics
VL  - 224
PB  - Elsevier
PY  - 2020
SN  - 1874-3919
ER  - 

TY  - JOUR
AB  - Proteasome dysfunction is emerging as a novel pathomechanism for the development of chronic obstructive pulmonary disease (COPD), a major leading cause of death in the world. Cigarette smoke, one of the main risk factors for COPD, impairs proteasome function in vitro and in vivo. In the present study, we dissected the molecular changes induced by cigarette smoke on the proteasome in lung epithelial cells and mouse lungs. 26S proteasome pull-down, MS interactome, and stoichiometry analyses indicated that 26S proteasome complexes become instable in cigarette smoke-treated lung epithelial cells as well as in lungs of mice after three day smoke exposure. The interactome of the 26S was clearly altered in mouse lungs upon smoke exposure but not in cells after 24 h of smoke exposure. Using native MS analysis of purified 20S proteasomes, we observed some destabilization of 20S complexes purified from cigarette smoke-exposed cells in the absence of any dominant and inhibitory modification of proteasomal proteins. Taken together, our results suggest that cigarette smoke induces minor but detectable changes in the stability of 20S and 26S proteasome complexes which might contribute to imbalanced proteostasis in a chronic setting as observed in chronic lung diseases associated with cigarette smoking.
AU  - Kammerl, I.E.
AU  - Caniard, A.
AU  - Merl-Pham, J.
AU  - Ben-Nissan, G.*
AU  - Mayr, C.H.
AU  - Mossina, A.
AU  - Geerlof, A.
AU  - Eickelberg, O.
AU  - Hauck, S.M.
AU  - Sharon, M.*
AU  - Meiners, S.
C1  - 54987
C2  - 46041
CY  - Po Box 211, 1000 Ae Amsterdam, Netherlands
SP  - 1-9
TI  - Dissecting the molecular effects of cigarette smoke on proteasome function.
JO  - J. Proteomics
VL  - 193
PB  - Elsevier Science Bv
PY  - 2019
SN  - 1874-3919
ER  - 

TY  - JOUR
AB  - The membrane protein expression repertoire of cells changes in course of activation. In equine recurrent uveitis (ERU), a spontaneous autoimmune disease in horses with relapsing and ultimately blinding inner eye inflammation, CD4 + T lymphocytes are the crucial pathogenic cells activated in the periphery directly prior to an inflammatory episode. In order to find relevant changes in the membrane proteome associated to disease, we sorted CD4 + lymphocytes and compared protein abundance from the generated proteome datasets of both healthy horses and ERU cases. We detected formin like 1, a key player in actin dependent cellular processes such as phagocytosis, cell adhesion and cell migration, with significantly higher abundance in the CD4 + cell membrane proteome of horses with ERU. In transmigration experiments, we demonstrated higher migration rate of cells originating from diseased animals connecting formin like 1 to the migratory ability of cells. These findings are the first description of formin like 1 in association to processes involved in migration of inflammatory CD4 + T cells across the blood-retinal barrier in a spontaneous ocular autoimmune disease and suggest formin like 1 to play a role in the molecular mechanisms of ERU disease pathogenesis. Data are available via ProteomeXchange with identifier PXD005384. Biological significance This comparative proteomic study of membrane proteins of CD4 + T cells identified a novel protein, formin like 1, with expression on the plasma cell membrane of equine CD4 + T cells and a significant change of abundance on CD4 + T cells of horses with a spontaneous autoimmune disease. Functional studies in a cell culture model for transmigration at the blood-retinal barrier (BRB) unraveled a strong impact of formin like 1 on migratory processes of CD4 + T cells across the BRB, a key event in uveitis pathogenesis. These findings provide novel knowledge about changes in the CD4 + immune cell membrane proteome in a spontaneously and naturally occurring autoimmune disease in horses with high relevance for veterinary medicine. Additionally, this model has proven translational quality for human medicine and provides novel proteomic information on autoimmune uveitis in man.
AU  - Degroote, R.L.*
AU  - Uhl, P.B.*
AU  - Amann, B.*
AU  - Krackhardt, A.M.*
AU  - Ueffing, M.
AU  - Hauck, S.M.
AU  - Deeg, C.A.*
C1  - 50299
C2  - 42395
CY  - Amsterdam
SP  - 102-108
TI  - Formin like 1 expression is increased on CD4 + T lymphocytes in spontaneous autoimmune uveitis.
JO  - J. Proteomics
VL  - 154
PB  - Elsevier Science Bv
PY  - 2017
SN  - 1874-3919
ER  - 

TY  - JOUR
AB  - Estrogens are suggested to lower the risk of developing metabolic syndrome in both sexes. In this study, we investigated how the increased circulating estrogen-to-androgen ratio (E/A) alters liver lipid metabolism in males. The cytochrome P450 aromatase (P450arom) is an enzyme converting androgens to estrogens. Male mice overexpressing human aromatase enzyme (AROM+ mice), and thus have high circulating E/A, were used as a model in this study. Proteomics and gene expression analyses indicated an increase in the peroxisomal β-oxidation in the liver of AROM+ mice as compared with their wild type littermates. Correspondingly, metabolomic analysis revealed a decrease in the amount of phosphatidylcholines with long-chain fatty acids in the plasma. With interest we noted that the expression of Cyp4a12a enzyme, which specifically metabolizes arachidonic acid (AA) to 20-hydroxy AA, was dramatically decreased in the AROM+ liver. As a consequence, increased amounts of phospholipids having AA as a fatty acid tail were detected in the plasma of the AROM+ mice. Overall, these observations demonstrate that high circulating E/A in males is linked to indicators of higher peroxisomal β-oxidation and lower AA metabolism in the liver. Furthermore, the plasma phospholipid profile reflects the changes in the liver lipid metabolism. BIOLOGICAL SIGNIFICANCE: The role of sex steroid hormones in the development of metabolic diseases is a topical issue. Lipid metabolism in both sexes supposedly benefits from estrogens, and low circulating estrogen to androgen ratio has been shown to lead to liver steatosis in males. However, there are no comprehensive studies showing the effects of sex steroid hormones on the expression of genes regulating liver lipid metabolism on both mRNA and protein levels. In this study a combination of quantitative MS-based proteome measurements and mRNA microarray both consistently indicated a set of genes that are deregulated in the liver of male mice having high circulating E/A. Interestingly, the results of targeted profiling of phospholipids in the plasma by LC-MS/MS were in line with the mRNA and protein measurements carried out in the liver, suggesting that plasma phospholipid profile could be used as an indicator of altered liver lipid metabolism.
AU  - Vehmas, A.P.*
AU  - Adam, M.*
AU  - Laajala, T.D.*
AU  - Kastenmüller, G.
AU  - Prehn, C.
AU  - Rozman, J.
AU  - Ohlsson, C.*
AU  - Fuchs, H.
AU  - Hrabě de Angelis, M.
AU  - Gailus-Durner, V.
AU  - Elo, L.L.*
AU  - Aittokallio, T.*
AU  - Adamski, J.
AU  - Corthals, G.L.*
AU  - Poutanen, M.*
AU  - Strauss, L.*
C1  - 47572
C2  - 38206
CY  - Amsterdam
SP  - 66-75
TI  - Liver lipid metabolism is altered by increased circulating estrogen to androgen ratio in male mouse.
JO  - J. Proteomics
VL  - 133
PB  - Elsevier Science Bv
PY  - 2016
SN  - 1874-3919
ER  - 

TY  - JOUR
AB  - Plasma membrane intrinsic proteins (PIPs) are one subfamily of aquaporins that mediate the transmembrane transport of water. To reveal their function in poplar, we generated transgenic poplar plants in which the translation of PIP genes was downregulated by RNA interference investigated these plants with a comprehensive leaf plasma membrane proteome and physiome analysis. First, inhibition of PIP synthesis strongly altered the leaf plasma membrane protein composition. Strikingly, several signaling components and transporters involved in the regulation of stomatal movement were differentially regulated in transgenic poplars. Furthermore, hormonal crosstalk related to abscisic acid, auxin and brassinosteroids was altered, in addition to cell wall biosynthesis/cutinization, the organization of cellular structures and membrane trafficking. A physiological analysis confirmed the proteomic results. The leaves had wider opened stomata and higher net CO2 assimilation and transpiration rates as well as greater mesophyll conductance for CO2 (gm) and leaf hydraulic conductance (Kleaf). Based on these results, we conclude that PIP proteins not only play essential roles in whole leaf water and CO2 flux but have important roles in the regulation of stomatal movement. Biological significance The present work is a comprehensive survey combining leaf plasma membrane proteomics and physiological methods to assess the functionality of the whole PIP subfamily in tree model system.
AU  - Bi, Z.
AU  - Merl-Pham, J.
AU  - Uehlein, N.*
AU  - Zimmer, I.
AU  - Mühlhans, S.
AU  - Aichler, M.
AU  - Walch, A.K.
AU  - Kaldenhoff, R.*
AU  - Palme, K.*
AU  - Schnitzler, J.-P.
AU  - Block, K.
C1  - 46545
C2  - 37642
SP  - 321-332
TI  - RNAi-mediated downregulation of poplar Plasma membrane Intrinsic Proteins (PIPs) changes plasma membrane proteome composition and affects leaf physiology.
JO  - J. Proteomics
VL  - 128
PY  - 2015
SN  - 1874-3919
ER  - 

TY  - JOUR
AB  - Age-related changes in vascular functioning are a harbinger of cardiovascular disease but the biological mechanisms during the progression of endothelial senescence have not been studied. We investigated alterations in the proteome and miRNA profiles in the course of replicative senescence using primary human umbilical vein endothelial cells as an in vitro vascular model. Quantitative proteomic profiling from early growth stage to senescence was performed by isotope-coded protein label coupled to LC-ESI-MS/MS analysis. Some proteins consistently changed their expression during the senescence whereas others appeared as deregulated only during the late senescence. The latter was accompanied by alterations in morphology of senescent endothelial cells. MicroRNA expression profiling revealed transient changes in the level of miR-16-5p, miR-28-3p and miR-886-5p in the early senescence, decrease in the level of miR-106b-3p at the late stage, and continuous changes in the expression of miR-181a-5p and miR-376a-3p during the whole senescence process. Integrating data on proteomic and microRNA changes indicated potential crosstalk between specific proteins and non-coding RNAs in the regulation of metabolism, cell cycle progression and cytoskeletal organization in the endothelial senescence. The knowledge of molecular targets that change during the senescence can ultimately contribute to a better understanding and prevention of age-related vascular diseases. SIGNIFICANCE: Endothelial cell senescence has been associated to age-related cardiovascular disease. Although replicative senescence was discovered more than 50years ago, detailed molecular mechanisms behind this phenomenon remain to be elucidated. This study describes detailed proteome and miRNAome analyses in the progressing senescence of human umbilical vein endothelial cells. The in silico integration of the two data sets indicated the important role of miRNAs in the regulation of cell cycle progression, adhesion and metabolic alteration in the cellular aging. This study adds to our knowledge of age-induced vascular alterations and contributes to our understanding and prevention of age-related vascular diseases.
AU  - Yentrapalli, R.
AU  - Azimzadeh, O.
AU  - Kraemer, A.
AU  - Malinowsky, K.*
AU  - Sarioglu, H.
AU  - Becker, K.F.*
AU  - Atkinson, M.J.
AU  - Mörtl, S.
AU  - Tapio, S.
C1  - 45035
C2  - 37146
CY  - Amsterdam
SP  - 12-23
TI  - Quantitative and integrated proteome and microRNA analysis of endothelial replicative senescence.
JO  - J. Proteomics
VL  - 126
PB  - Elsevier Science Bv
PY  - 2015
SN  - 1874-3919
ER  - 

TY  - JOUR
AB  - In the present study, we performed a large-scale protein analysis based on 2-DE DIGE to examine the effects of ozone on the leaves of juvenile European beech (Fagus sylvatica L.), one of the most important deciduous trees in central Europe. To this end, beech trees were grown under field conditions and subjected to ambient and twice ambient ozone concentrations during the vegetation periods of four consecutive years. The twice ambient ozone concentration altered the abundance of 237 protein spots, which showed relative ratios higher than 30 % compared to the ambient control trees. A total of 74 protein spots were subjected to mass spectrometry identification (LC-MS/MS), followed by homology-driven searches. The differentially expressed proteins participate in key biological processes including the Calvin cycle and photosynthesis, carbon metabolism, defense- and stress-related responses, detoxification mechanisms, protein folding and degradation, and mechanisms involved in senescence. The ozone-induced responses provide evidence of a changing carbon metabolism and counteraction against increased levels of reactive oxygen species. Biological Significance This study provides useful information on how European beech, an economically and ecologically important tree, reacts on the molecular level to increased ozone concentrations expected in the near future. The main emphasis in the present study was placed on identifying differentially abundant proteins after long-term ozone exposure under climatically realistic settings, rather than short-term responses or reactions under laboratory conditions. Additionally, using nursery-grown beech trees, we took into account the natural genotypic variation of this species. As such, the results presented here provide information on molecular responses to ozone in an experimental plant system at very close to natural conditions. Furthermore, this proteomic approach was supported by previous studies on the present experiment. Ultimately, the combination of this proteomic approach with several approaches including transcriptomics, analysis of non-structural carbohydrates, and morphological effects contributes to a more global picture of how beech trees react under increased ozone concentrations.
AU  - Kerner, R.C.
AU  - Delgado-Eckert, E.*
AU  - Ernst, D.
AU  - Dupuy, J.W.*
AU  - Thorsten Grams, E.E.*
AU  - Winkler, J.B.
AU  - Lindermayr, C.
AU  - Müller-Starck, G.*
C1  - 31551
C2  - 34565
SP  - 417-435
TI  - Large-scale protein analysis of European beech trees following four vegetation periods of twice ambient ozone exposure.
JO  - J. Proteomics
VL  - 109
PY  - 2014
SN  - 1874-3919
ER  - 

TY  - JOUR
AB  - Retinal pigment epithelium (RPE) builds the outer blood-retinal barrier of the eye and plays an important role in pathogenesis of the sight threatening disease equine recurrent uveitis (ERU). ERU is a spontaneous autoimmune mediated inflammatory disease characterised by the breakdown of the outer blood-retinal barrier and an influx of autoaggressive T-cells into the inner eye. Therefore, identification of molecular mechanisms contributing to changed function of blood-retinal barrier in ERU is important for understanding of pathophysiology. Cell surface proteins of RPE collected from healthy horses and horses with ERU were captured by in situ biotinylation and analysed with high resolution mass spectrometry coupled to liquid chromatography (LC-MS/MS) to identify differentially expressed proteins. With label free differential proteomics, a total of 27 differently expressed cell surface proteins in diseased RPE could be detected. Significant down-regulation of three very interesting proteins, synaptotagmin 1, basigin and collectrin was verified and further characterised. BIOLOGICAL SIGNIFICANCE: We applied an innovative and successful method to detect changes in the plasma cell surface proteome of RPE cells in a spontaneous inflammatory eye disease, serving as a valuable model for human autoimmune uveitis. We were able to identify 27 differentially expressed plasma cell membrane proteins, including synaptotagmin 1, basigin and collectrin, which play important roles in cell adhesion, transport and cell communication.
AU  - Uhl, P.B.*
AU  - Szober, C.M.*
AU  - Amann, B.*
AU  - Alge-Priglinger, C.S.*
AU  - Ueffing, M.
AU  - Hauck, S.M.
AU  - Deeg, C.A.*
C1  - 31739
C2  - 34705
SP  - 50-62
TI  - In situ cell surface proteomics reveals differentially expressed membrane proteins in retinal pigment epithelial cells during autoimmune uveitis.
JO  - J. Proteomics
VL  - 109
PY  - 2014
SN  - 1874-3919
ER  - 

TY  - JOUR
AB  - Formin-like 1 (FMNL1) is a formin-related protein highly expressed in hematopoietic cells and overexpressed in leukemias as well as diverse transformed cell lines. It has been described to play a role in diverse functions of hematopoietic cells such as phagocytosis of macrophages as well as polarization and cytotoxicity of T cells. However, the specific role of FMNL1 in these processes has not been clarified yet and regulation by interaction partners in primary hematopoietic cells has never been investigated. We performed a proteomic screen for investigation of the interactome of FMNL1 in primary hematopoietic cells resulting in the identification of a number of interaction partners. Bioinformatic analysis considering semantic similarity suggested the giant protein AHNAK1 to be an essential interaction partner of FMNL1. We confirmed AHNAK1 as a general binding partner for FMNL1 in diverse hematopoietic cells and demonstrate that the N-terminal part of FMNL1 binds to the C-terminus of AHNAK1. Moreover, we show that the constitutively activated form of FMNL1 (FMNL1γ) induces localization of AHNAK1 to the cell membrane. Finally, we provide evidence that overexpression or knock down of FMNL1 has an impact on the capacitative calcium influx after ionomycin-mediated activation of diverse cell lines and primary cells.
AU  - Han, Y.*
AU  - Yu, G.*
AU  - Sarioglu, H.
AU  - Caballero-Martinez, A.*
AU  - Schlott, F.*
AU  - Ueffing, M.
AU  - Haase, H.*
AU  - Peschel, C.*
AU  - Krackhardt, A.M.
C1  - 11676
C2  - 30746
SP  - 72-82
TI  - Proteomic investigation of the interactome of FMNL1 in hematopoietic cells unveils a role in calcium-dependent membrane plasticity.
JO  - J. Proteomics
VL  - 78
PB  - Elsevier
PY  - 2013
SN  - 1874-3919
ER  - 

TY  - JOUR
AB  - UNLABELLED: Among the idiopathic interstitial pneumonias (IIP), the two entities IPF and NSIP seem to be clinically related, but NSIP has a better outcome. The proteomic signatures which distinguish NSIP from IPF remain still elusive. We therefore performed comparative proteomic analysis of peripheral lung tissue from patients with sporadic IPF (n=14) and fibrotic NSIP (fNSIP, n=8) and organ donors (Controls, n=10), by using the 2-dimensional DIGE technique and MALDI-TOF-MS. The study revealed that the proteomic profiles of IPF and fNSIP were quite similar. Among the upregulated proteins in IPF and fNSIP were stress-induced genes involved in the ER stress-pathway, whereas downregulated proteins in IPF and fNSIP included antiapoptotic factors and antifibrotic molecules. The comparison fNSIP versus IPF indicated upregulation of subunits of the proteasome activator complex and antioxidant enzymes of the peroxiredoxin family. We conclude, that only few protein expression changes exist between IPF and fNSIP, and that epithelial ER- and oxidative stress play a major role in the pathogenesis of both diseases. In contrast to IPF, intracellular clearance of ROS and misfolded protein carbonyls seem to be enhanced in fNSIP due to enhanced expression of antioxidant acting proteins, and may explain the better outcome and survival in patients with fNSIP. BIOLOGICAL SIGNIFICANCE: IPF and fibrotic NSIP (fNSIP) belong to the idiopathic interstitial pneumonias and are usually fatal, but fNSIP has a better outcome. In order to identify molecular mechanisms and differences between IPF and fNSIP, we herein present results of a comparative proteome analysis of IPF, fNSIP and control lung tissue. Our data including validation experiments suggest that ER stress and a general stress-response as well as the decline of antioxidant capacity in alveolar epithelium is key in the pathogenesis of IPF and fNSIP. In addition, we could observe a signature of an increased alveolar epithelial protection against oxidative and ER-stress in fNSIP as compared to IPF, which could help to explain the better outcome of fNSIP patients.
AU  - Korfei, M.*
AU  - von der Beck, D.*
AU  - Henneke, I.*
AU  - Markart, P.*
AU  - Ruppert, C.*
AU  - Mahavadi, P.*
AU  - Ghanim, B.*
AU  - Klepetko, W.*
AU  - Fink, L.*
AU  - Meiners, S.
AU  - Krämer, O.H.*
AU  - Seeger, W.*
AU  - Vancheri, C.*
AU  - Guenther, A.*
C1  - 27962
C2  - 32879
SP  - 109-128
TI  - Comparative proteome analysis of lung tissue from patients with idiopathic pulmonary fibrosis (IPF), non-specific interstitial pneumonia (NSIP) and organ donors.
JO  - J. Proteomics
VL  - 85
PB  - Elsevier Science
PY  - 2013
SN  - 1874-3919
ER  - 

TY  - JOUR
AB  - MALDI imaging mass spectrometry (MALDI-imaging) has emerged as a spatially-resolved label-free bioanalytical technique for direct analysis of biological samples and was recently introduced for analysis of 3D tissue specimens. We present a new experimental and computational pipeline for molecular analysis of tissue specimens which integrates 3D MALDI-imaging, magnetic resonance imaging (MRI), and histological staining and microscopy, and evaluate the pipeline by applying it to analysis of a mouse kidney. To ensure sample integrity and reproducible sectioning, we utilized the PAXgene fixation and paraffin embedding and proved its compatibility with MRI. Altogether, 122 serial sections of the kidney were analyzed using MALDI-imaging, resulting in a 3D dataset of 200 GB comprised of 2 million spectra. We show that elastic image registration better compensates for local distortions of tissue sections. The computational analysis of 3D MALDI-imaging data was performed using our spatial segmentation pipeline which determines regions of distinct molecular composition and finds m/z-values co-localized with these regions. For facilitated interpretation of 3D distribution of ions, we evaluated isosurfaces providing simplified visualization. We present the data in a multimodal fashion combining 3D MALDI-imaging with the MRI volume rendering and with light microscopic images of histologically stained sections. BIOLOGICAL SIGNIFICANCE: Our novel experimental and computational pipeline for 3D MALDI-imaging can be applied to address clinical questions such as proteomic analysis of the tumor morphologic heterogeneity. Examining the protein distribution as well as the drug distribution throughout an entire tumor using our pipeline will facilitate understanding of the molecular mechanisms of carcinogenesis.
AU  - Oetjen, J.*
AU  - Aichler, M.
AU  - Trede, D.*
AU  - Strehlow, J.*
AU  - Berger, J.*
AU  - Heldmann, S.*
AU  - Becker, M.*
AU  - Gottschalk, M.*
AU  - Kobarg, J.H.*
AU  - Wirtz, S.*
AU  - Schiffler, S.*
AU  - Thiele, H.*
AU  - Walch, A.K.
AU  - Maass, P.*
AU  - Alexandrov, T.*
C1  - 23727
C2  - 31259
SP  - 52-60
TI  - MRI-compatible pipeline for three-dimensional MALDI imaging mass spectrometry using PAXgene fixation.
JO  - J. Proteomics
VL  - 90
PB  - Elsevier Science
PY  - 2013
SN  - 1874-3919
ER  - 

TY  - JOUR
AB  - [object Object]: New biomarkers are needed to improve the specificity of prostate cancer detection and characterisation of individual tumors. In a proteomics profiling approach using MALDI-MS tissue imaging on frozen tissue sections, we identified discriminating masses. Imaging analysis of cancer, non-malignant benign epithelium and stromal areas of 15 prostatectomy specimens in a test and 10 in a validation set identified characteristic m/z peaks for each tissue type, e.g. m/z 10775 for benign epithelial, m/z 6284 and m/z 6657.5 for cancer and m/z 4965 for stromal tissue. A 10-fold cross-validation analysis showed highest discriminatory ability to separate tissue types for m/z 6284 and m/z 6657.5, both overexpressed in cancer, and a multicomponent mass peak cluster at m/z 10775-10797.4 overexpressed in benign epithelial tissue. ROC AUC values for these three masses ranged from 0.85 to 0.95 in the discrimination of malignant and non-malignant tissue. To identify the underlying proteins, prostate whole tissue extract was separated by nano-HPLC and subjected to MALDI TOF/TOF analysis. Proteins in fractions containing discriminatory m/z masses were identified by MS/MS analysis and candidate marker proteins subsequently validated by immunohistochemistry (IHC). Biliverdin reductase B (BLVRB) turned out to be overexpressed in PCa tissue. BIOLOGICAL SIGNIFICANCE: In this study on cryosections of radical prostatectomies of prostate cancer patients, we performed a MALDI-MS tissue imaging analysis and a consecutive protein identification of significant m/z masses by nano-HPLC, MALDI TOF/TOF and MS/MS analysis. We identified BLVRB as a potential biomarker in the discrimination of PCa and benign tissue, also suggesting BVR as a feasible therapeutic target.
AU  - Pallua, J.D.*
AU  - Schaefer, G.*
AU  - Seifarth, C.*
AU  - Becker, M.*
AU  - Meding, S.
AU  - Rauser, S.
AU  - Walch, A.K.
AU  - Handler, M.*
AU  - Netzer, M.*
AU  - Popovscaia, M.*
AU  - Osl, M.*
AU  - Baumgartner, C.*
AU  - Lindner, H.*
AU  - Kremser, L.*
AU  - Sarg, B.*
AU  - Bartsch, G.*
AU  - Huck, C.W.*
AU  - Bonn, G.K.*
AU  - Klocker, H.*
C1  - 27042
C2  - 32495
SP  - 500-514
TI  - MALDI-MS tissue imaging identification of biliverdin reductase B overexpression in prostate cancer.
JO  - J. Proteomics
VL  - 91
IS  - 8
PB  - Elsevier Science
PY  - 2013
SN  - 1874-3919
ER  - 

TY  - JOUR
AB  - Qualitative proteome profiling of formalin-fixed, paraffin-embedded (FFPE) tissue is advancing the field of clinical proteomics. However, quantitative proteome analysis of FFPE tissue is hampered by the lack of an efficient labelling method. The usage of conventional protein labelling on FFPE tissue has turned out to be inefficient. Classical labelling targets lysine residues that are blocked by the formalin treatment. The aim of this study was to establish a quantitative proteomics analysis of FFPE tissue by combining the label-free approach with optimised protein extraction and separation conditions. As a model system we used FFPE heart tissue of control and exposed C57BL/6 mice after total body irradiation using a gamma ray dose of 3 gray. We identified 32 deregulated proteins (p≤0.05) in irradiated hearts 24h after the exposure. The proteomics data were further evaluated and validated by bioinformatics and immunoblotting investigation. In good agreement with our previous results using fresh-frozen tissue, the analysis indicated radiation-induced alterations in three main biological pathways: respiratory chain, lipid metabolism and pyruvate metabolism. The label-free approach enables the quantitative measurement of radiation-induced alterations in FFPE tissue and facilitates retrospective biomarker identification using clinical archives.
AU  - Azimzadeh, O.
AU  - Scherthan, H.*
AU  - Yentrapalli, R.
AU  - Barjaktarovic, Z.
AU  - Ueffing, M.*
AU  - Conrad, M.
AU  - Neff, F.
AU  - Calzada-Wack, J.
AU  - Aubele, M.
AU  - Buske, C.*
AU  - Atkinson, M.J.
AU  - Hauck, S.M.*
AU  - Tapio, S.
C1  - 7390
C2  - 29697
SP  - 2384-2395
TI  - Label-free protein profiling of formalin-fixed paraffin-embedded (FFPE) heart tissue reveals immediate mitochondrial impairment after ionising radiation.
JO  - J. Proteomics
VL  - 75
IS  - 8
PB  - Elsevier
PY  - 2012
SN  - 1874-3919
ER  - 

TY  - JOUR
AB  - The molecular mechanism which enables activated immune cells to cross the blood-retinal barrier in spontaneous autoimmune uveitis is yet to be unraveled. Equine recurrent uveitis is the only spontaneous animal model allowing us to investigate the autoimmune mediated transformation of leukocytes in the course of this sight threatening disease. Hypothesizing that peripheral blood immune cells change their protein expression pattern in spontaneous autoimmune uveitis, we used DIGE to detect proteins with altered abundance comparing peripheral immune cells of healthy and ERU diseased horses. Among others, we found a significant downregulation of talin 1 in peripheral blood granulocytes of ERU specimen, pointing to changes in beta integrin activation and indicating a significant role of the innate immune system in spontaneous autoimmune diseases. This article is part of a Special Issue entitled: SI: Farm animal proteomics.
AU  - Degroote, R.L.*
AU  - Hauck, S.M.
AU  - Kremmer, E.
AU  - Amann, B.*
AU  - Ueffing, M.
AU  - Deeg, C.A.*
C1  - 8580
C2  - 30278
SP  - 4536-4544
TI  - Altered expression of talin 1 in peripheral immune cells points to a significant role of the innate immune system in spontaneous autoimmune uveitis.
JO  - J. Proteomics
VL  - 75
IS  - 14
PB  - Elsevier
PY  - 2012
SN  - 1874-3919
ER  - 

TY  - JOUR
AB  - To characterize proteomic changes found in Barrett's adenocarcinoma and its premalignant stages, the proteomic profiles of histologically defined precursor and invasive carcinoma lesions were analyzed by MALDI imaging MS. For a primary proteomic screening, a discovery cohort of 38 fresh frozen Barrett's adenocarcinoma patient tissue samples was used. The goal was to find proteins that might be used as markers for monitoring cancer development as well as for predicting regional lymph node metastasis and disease outcome. Using mass spectrometry for protein identification and validating the results by immunohistochemistry on an independent validation set, we could identify two of 60 differentially expressed m/z species between Barrett's adenocarcinoma and the precursor lesion: COX7A2 and S100-A10. Furthermore, among 22m/z species that are differentially expressed in Barrett's adenocarcinoma cases with and without regional lymph node metastasis, one was identified as TAGLN2. In the validation set, we found a correlation of the expression levels of COX7A2 and TAGLN2 with a poor prognosis while S100-A10 was confirmed by multivariate analysis as a novel independent prognostic factor in Barrett's adenocarcinoma. Our results underscore the high potential of MALDI imaging for revealing new biologically significant molecular details from cancer tissues which might have potential for clinical application. This article is part of a Special Issue entitled: Translational Proteomics.
AU  - Elsner, M.
AU  - Rauser, S.
AU  - Maier, S.K.
AU  - Schöne, C.
AU  - Balluff, B.
AU  - Meding, S.
AU  - Jung, G.
AU  - Nipp, M.
AU  - Sarioglu, H.
AU  - Maccarrone, G.*
AU  - Aichler, M.
AU  - Feuchtinger, A.
AU  - Langer, R.*
AU  - Jütting, U.
AU  - Feith, M.*
AU  - Kuster, B.*
AU  - Ueffing, M.
AU  - Zitzelsberger, H.
AU  - Höfler, H.
AU  - Walch, A.K.
C1  - 7383
C2  - 29690
SP  - 4693-4704
TI  - MALDI imaging mass spectrometry reveals COX7A2, TAGLN2 and S100-A10 as novel prognostic markers in Barrett's adenocarcinoma.
JO  - J. Proteomics
VL  - 75
IS  - 15
PB  - Elsevier
PY  - 2012
SN  - 1874-3919
ER  - 

TY  - JOUR
AB  - Equine recurrent uveitis is a severe and frequent blinding disease in horses which presents with auto-reactive invading T-cells, resulting in the destruction of the inner eye. Infiltration of inflammatory cells into the retina and vitreous is driven by currently unknown guidance cues, however surgical removal of the vitreous (vitrectomy) has proven therapeutically successful. Therefore, proteomic analyses of vitrectomy samples are likely to result in detection of proteins contributing to disease pathogenesis. Vitreous from healthy and ERU diseased horses were directly compared by quantitative mass spectrometry based on label-free quantification of peak intensities across samples. We found a significant upregulation of complement and coagulation cascades and downregulation of negative paracrine regulators of canonical Wnt signalling including the Wnt signalling inhibitors DKK3 and SFRP2. Based on immunohistochemistry, both proteins are expressed in equine retina and suggest localisation to retinal Muller glial cells (RMG), which may be the source cells for these proteins. Furthermore, retinal expression levels and patterns of DKK3 change in response to ERU. Since many other regulated proteins identified here are associated with RMG cells, these cells qualify as the prime responders to autoimmune triggers. This article is part of a Special Issue entitled: "Farm animal proteomics".
AU  - Hauck, S.M.
AU  - Hofmaier, F.*
AU  - Dietter, J.
AU  - Swadzba, M.E.*
AU  - Blindert, M.
AU  - Amann, B.*
AU  - Behler, J.
AU  - Kremmer, E.
AU  - Ueffing, M.
AU  - Deeg, C.A.*
C1  - 8581
C2  - 30279
SP  - 4545-4554
TI  - Label-free LC-MSMS analysis of vitreous from autoimmune uveitis reveals a significant decrease in secreted Wnt signalling inhibitors DKK3 and SFRP2.
JO  - J. Proteomics
VL  - 75
IS  - 14
PB  - Elsevier
PY  - 2012
SN  - 1874-3919
ER  - 

TY  - JOUR
AB  - Cenococcum geophilum is a widely distributed ectomycorrhizal fungus potentially playing a significant role in resistance and resilience mechanisms of its tree hosts exposed to drought stress. In this study, we performed a large scale protein analysis in pure cultures of C. geophilum in order to gain first global insights into the proteome assembly of this fungus. Using 1-D gel electrophoresis coupled with ESI-MS/MS, we indentified 638 unique proteins. Most of these proteins were related to the metabolic and cellular processes, and the transport machinery of cells. In a second step, we examined the influence of water deprivation on the proteome of C. geophilum pure cultures at three time points of gradually imposed drought. The results indicated that 12 proteins were differentially abundant in mycelia subjected to drought compared to controls. The induced responses in C. geophilum point towards regulation of osmotic stress, maintainance of cell integrity, and counteracting increased levels of reactive oxygen species formed during water deprivation.
AU  - Kerner, R.
AU  - Delgado-Eckert, E.*
AU  - Del Castillo, E.*
AU  - Müller-Starck, G.*
AU  - Peter, M.*
AU  - Kuster, B.*
AU  - Tisserant, E.*
AU  - Pritsch, K.
C1  - 8165
C2  - 30006
SP  - 3707-3719
TI  - Comprehensive proteome analysis in Cenococcum geophilum Fr. as a tool to discover drought-related proteins.
JO  - J. Proteomics
VL  - 75
IS  - 12
PB  - Elsevier
PY  - 2012
SN  - 1874-3919
ER  - 

TY  - JOUR
AB  - Epidemiological data show that ionising radiation increases the risk of cardiovascular disease. The endothelium is one of the main targets of radiation-induced damage. Rapid radiation-induced alterations in the biological processes were investigated after exposure to a clinically relevant radiation dose (2.5Gy gamma radiation). The changes in protein expression were determined using the human endothelial cell line EA.hy926 as a model. Two complementary proteomic approaches, SILAC (Stable Isotope Labelling with Amino acids in Cell culture) and 2D-DIGE (Two Dimensional Difference-in-Gel-Electrophoresis) were used. The proteomes of the endothelial cells were analysed 4h and 24h after irradiation. Differentially expressed proteins were identified and quantified by MALDI-TOF/TOF and LTQ Orbitrap tandem mass spectrometry. The deregulated proteins were mainly categorised in four key pathways: (i) glycolysis/gluconeogenesis and synthesis/degradation of ketone bodies, (ii) oxidative phosphorylation, (iii) Rho-mediated cell motility and (iv) non-homologous end joining. We suggest that these alterations facilitate the repair processes needed to overcome the stress caused by irradiation and are indicative of the vascular damage leading to radiation-induced cardio- and cerebrovascular impairment.
AU  - Sriharshan, A.
AU  - Boldt, K.
AU  - Sarioglu, H.
AU  - Barjaktarovic, Z.
AU  - Azimzadeh, O.
AU  - Hieber, L.
AU  - Zitzelsberger, H.
AU  - Ueffing, M.
AU  - Atkinson, M.J.
AU  - Tapio, S.
C1  - 7392
C2  - 29699
SP  - 2319-2330
TI  - Proteomic analysis by SILAC and 2D-DIGE reveals radiation-induced endothelial response: Four key pathways.
JO  - J. Proteomics
VL  - 75
IS  - 8
PB  - Elsevier
PY  - 2012
SN  - 1874-3919
ER  - 

TY  - JOUR
AB  - In the last decade, imaging mass spectrometry has seen incredible technological advances in its applications to biological samples. One computational method of data mining in this field is the spatial segmentation of a sample, which produces a segmentation map highlighting chemically similar regions. An important issue for any imaging mass spectrometry technology is its relatively low spatial or lateral resolution (i.e. a large size of pixel) as compared with microscopy. Thus, the spatial resolution of a segmentation map is also relatively low, that complicates its visual examination and interpretation when compared with microscopy data, as well as reduces the accuracy of any automated comparison. We address this issue by proposing an approach to improve the spatial resolution of a segmentation map. Given a segmentation map, our method magnifies it up to some factor, producing a super-resolution segmentation map. The super-resolution map can be overlaid and compared with a high-res microscopy image. The proposed method is based on recent advances in image processing and smoothes the "pixilated" region boundaries while preserving fine details. Moreover, it neither eliminates nor splits any region. We evaluated the proposed super-resolution segmentation approach on three MALDI-imaging datasets of human tissue sections and demonstrated the superiority of the super-segmentation maps over standard segmentation maps.
AU  - Alexandrov, T.*
AU  - Meding, S.
AU  - Trede, D.*
AU  - Kobarg, J.H.*
AU  - Balluff, B.
AU  - Walch, A.K.
AU  - Thiele, H.*
AU  - Maass, P.*
C1  - 6712
C2  - 29149
SP  - 237-245
TI  - Super-resolution segmentation of imaging mass spectrometry data: Solving the issue of low lateral resolution.
JO  - J. Proteomics
VL  - 75
IS  - 1
PB  - Elsevier
PY  - 2011
SN  - 1874-3919
ER  - 

TY  - JOUR
AB  - Proteome analyses suffer from the large complexity of even small proteomes. Additionally, in many protein samples a few highly abundant proteins are hindering detailed proteomic studies, since they mask low abundant proteins. Recently, a new technology has emerged, which reduces dynamic range of protein concentrations within a given sample using combinatorial hexapeptide ligand libraries (CPLLs). This technique has been widely used in the microbial, animal and human fields and is now going to enter plant research. It can be a useful tool for fractionation of protein samples and might help to get a deeper insight into specific plant proteomes. In this review we describe the CPLL protein fractionation, summarize its possible applications in the plant field and discuss the limitations of this method.
AU  - Fröhlich, A.
AU  - Lindermayr, C.
C1  - 4268
C2  - 28826
SP  - 1182-1189
TI  - Deep insights into the plant proteome by pretreatment with combinatorial hexapeptide ligand libraries.
JO  - J. Proteomics
VL  - 74
IS  - 8
PB  - Elsevier
PY  - 2011
SN  - 1874-3919
ER  - 

TY  - JOUR
AB  - This study highlights proteomic and enzymatic changes in roots and leaves of actively growing poplar plants upon a cadmium stress exposure. Proteomic changes in response to a short-term (14 days), as well as a longer term (56 days) treatment are observed between the different organs. In leaves, stress-related proteins, like heat shock proteins, proteinases and pathogenesis-related proteins increased in abundance. A response similar to a hypersensitive response upon plant-pathogen interaction seemed to be induced. Concerning roots it appeared that the metabolic impact of cadmium was more deleterious than in leaves. This is evidenced by the early increase in abundance of many typical stress-related proteins like heat shock proteins, or glutathione-S-transferases, while most proteins from the primary metabolism (glycolysis, tricarboxylic acid cycle, nitrogen metabolism, sulfur metabolism) were severely decreased in abundance. Additionally the impact of cadmium on the glutathione metabolism could be assessed by activity assays of several important enzymes. Cadmium treatment had an inhibitory effect on glutathione reductase and ascorbate peroxidase in leaves, but not in roots. Conversely, glutathione-S-transferase showed a higher activity (and abundance) in roots but not in leaves.
AU  - Kieffer, P.*
AU  - Schröder, P.
AU  - Dommes, J.*
AU  - Hoffmann, L.*
AU  - Renaut, J.*
AU  - Hausman, J.F.*
C1  - 1001
C2  - 27017
SP  - 379-396
TI  - Proteomic and enzymatic response of poplar to cadmium stress.
JO  - J. Proteomics
VL  - 72
IS  - 3
PB  - Elsevier Science Bv
PY  - 2009
SN  - 1874-3919
ER  - 

TY  - JOUR
AB  - During the last two decades nitric oxide (NO) has emerged as a new chemical messenger in plant biology, which is involved in many different physiological processes, such as plant defense, transpiration and gas exchange, seed germination, and root development. Protein S-nitrosylation, the post-translational modification of thiol residues, has been suggested to be the most important mechanism for transduction of the bioactivity of NO. The characterization of protein S-nitrosylation as well as the physiological relevance of this type of modification is essential information, which is necessary to understand the function of NO in plants. In this review we focus on the formation of nitrosothiols and describe the chemistry of NO and thiol groups. Furthermore, different methods for detection of S-nitrosothiols are highlighted and the function of S-nitrosylation in plants is discussed.
AU  - Lindermayr, C.
AU  - Durner, J.
C1  - 1451
C2  - 26933
SP  - 1-9
TI  - S-Nitrosylation in plants: Pattern and function.
JO  - J. Proteomics
VL  - 73
IS  - 1
PB  - ELSEVIER SCIENCE BV
PY  - 2009
SN  - 1874-3919
ER  - 

TY  - JOUR
AB  - Here we report on the effect of the bacterial elicitor harpin from Pseudomonas syringae on Arabidopsis thaliana with an emphasis on transcriptional profiling and on changes of the mitochondrial proteome. Interestingly, of the currently about 400 identified mitochondrial proteins, transcriptional profiling by genome-wide DNA-microarray analyses revealed a total of 192 genes that showed significant changes in transcript abundance in response to the bacterial elicitor. The most dramatic changes were observed for the mitochondrial protein import apparatus of which 70% of all genes were induced. Proteomic analyses by 2D-PAGE and MALDI-TOF Mass Spectrometry for peptide fingerprint analysis confirmed the corresponding changes within the mitochondrial proteome in 28 cases. Strikingly, we found an accumulation of virtually all members of the TCA cycle. In sum, our results point to an involvement of mitochondria in the response of plant cells to the harpin elicitor, which becomes apparent both at the transcriptomic as well as proteomic level.
AU  - Livaja, M.
AU  - Palmieri, M.C.
AU  - von Rad, U.
AU  - Durner, J.
C1  - 381
C2  - 25358
SP  - 148-159
TI  - The effect of the bacterial effector protein harpin on transcriptional profile and mitochondrial proteins of Arabidopsis thaliana.
JO  - J. Proteomics
VL  - 71
IS  - 2
PB  - Elsevier
PY  - 2008
SN  - 1874-3919
ER  -