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Müller, G. ; Herling, A.W.* ; Stemmer, K. ; Lechner, A. ; Tschöp, M.H.

Chip-based sensing for release of unprocessed cell surface proteins in vitro and in serum and its (patho)physiological relevance.

Am. J. Physiol. Endocrinol. Metab. 317, https://doi.org/ 10.6084/m9.figshare.7994312.v1, E212-E233 (2019)
Verlagsversion Postprint Forschungsdaten DOI PMC
Open Access Gold
To study the possibility that certain components of eukaryotic plasma membranes are released under certain (patho)physiological conditions, a chip-based sensor was developed for the detection of cell surface proteins, which are anchored at the outer leaflet of eukaryotic plasma membranes by a covalently attached glycolipid, exclusively, and might be prone to spontaneous or regulated release on the basis of their amphiphilic character. For this, unprocessed, full-length glycosylphosphatidylinositol-anchored proteins (GPI-AP), together with associated phospholipids, were specifically captured and detected by a chip- and microfluidic channel-based sensor, leading to changes in phase and amplitude of surface acoustic waves (SAW) propagating over the chip surface. Unprocessed GPI-AP in complex with lipids were found to be released from rat adipocyte plasma membranes immobilized on the chip, which was dependent on the flow rate and composition of the buffer stream. The complexes were identified in the incubation medium of primary rat adipocytes, in correlation to the cell size, and in rat as well as human serum. With rats, the measured changes in SAW phase shift, reflecting specific mass/size or amount of the unprocessed GPI-AP in complex with lipids, and SAW amplitude, reflecting their viscoelasticity, enabled the differentiation between the lean and obese (high-fat diet) state, and the normal (Wistar) and hyperinsulinemic (Zucker fatty) as well as hyperinsulinemic hyperglycemic (Zucker diabetic fatty) state. Thus chip-based sensing for complexes of unprocessed GPI-AP and lipids reveals the inherently labile anchorage of GPI-AP at plasma membranes and their susceptibility for release in response to (intrinsic/extrinsic) cues of metabolic relevance and may, therefore, be useful for monitoring of (pre-)diabetic disease states.
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Publikationstyp Artikel: Journalartikel
Dokumenttyp Wissenschaftlicher Artikel
Schlagwörter Adipocytes ; Biosensor ; Diabetes ; Glycolipid-anchored Cell Surface Proteins ; Metabolic Stress ; Obesity; Gpi-anchored Proteins; Acoustic-wave; Lipid Rafts; Phospholipase-c; Binding; Model; Acetylcholinesterase; Phosphoinositolglycans; Biomarkers; Complex
Sprache englisch
Veröffentlichungsjahr 2019
HGF-Berichtsjahr 2019
ISSN (print) / ISBN 0193-1849
e-ISSN 1522-1555
Zeitschrift American Journal of Physiology - Endocrinology and Metabolism
Quellenangaben Band: 317, Heft: 2, Seiten: E212-E233, Artikelnummer: , Supplement: https://doi.org/ 10.6084/m9.figshare.7994312.v1
Verlag American Physiological Society
Verlagsort 9650 Rockville Pike, Bethesda, Md 20814 Usa
Begutachtungsstatus Peer reviewed
Institut(e) Institute of Diabetes and Obesity (IDO)
Institute of Experimental Genetics (IEG)
POF Topic(s) 30201 - Metabolic Health
90000 - German Center for Diabetes Research
Forschungsfeld(er) Helmholtz Diabetes Center
Genetics and Epidemiology
PSP-Element(e) G-502200-001
G-501900-221
G-521500-002
DOI 10.1152/ajpendo.00079.2019
WOS ID WOS:000475373900004
Scopus ID 85069799889
PubMed ID 31039006
Erfassungsdatum 2019-05-13
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