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Neukam, M. ; Sala, P. ; Brunner, A.D.* ; Ganß, K. ; Palladini, A. ; Grzybek, M. ; Topcheva, O. ; Vasiljevic, J. ; Broichhagen, J.* ; Johnsson, K.* ; Kurth, T.* ; Mann, M.* ; Coskun, Ü. ; Solimena, M.

Purification of time-resolved insulin granules reveals proteomic and lipidomic changes during granule aging.

Cell Rep. 43:113836 (2024)
Publ. Version/Full Text DOI PMC
Open Access Gold
Creative Commons Lizenzvertrag
Endocrine cells employ regulated exocytosis of secretory granules to secrete hormones and neurotransmitters. Secretory granule exocytosis depends on spatiotemporal variables such as proximity to the plasma membrane and age, with newly generated granules being preferentially released. Despite recent advances, we lack a comprehensive view of the molecular composition of insulin granules and associated changes over their lifetime. Here, we report a strategy for the purification of insulin secretory granules of distinct age from insulinoma INS-1 cells. Tagging the granule-resident protein phogrin with a cleavable CLIP tag, we obtain intact fractions of age-distinct granules for proteomic and lipidomic analyses. We find that the lipid composition changes over time, along with the physical properties of the membrane, and that kinesin-1 heavy chain (KIF5b) as well as Ras-related protein 3a (RAB3a) associate preferentially with younger granules. Further, we identify the Rho GTPase-activating protein (ARHGAP1) as a cytosolic factor associated with insulin granules.
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Publication type Article: Journal article
Document type Scientific Article
Keywords Arhgap1 ; Clip Tag ; Cp: Cell Biology ; Cp: Metabolism ; Kif5b ; Rab3a ; Snap Tag ; Granule Aging ; Insulin Granule ; Lipidomics ; Organelle Purification ; Proteomics; Secretory Granules; Preferential Release; Phosphatidic-acid; Beta-cell; C-laurdan; Protein; Glucose; Vesicles; Cholesterol; Trafficking
Language english
Publication Year 2024
HGF-reported in Year 2024
ISSN (print) / ISBN 2211-1247
e-ISSN 2211-1247
Journal Cell Reports
Quellenangaben Volume: 43, Issue: 3, Pages: , Article Number: 113836 Supplement: ,
Publisher Cell Press
Publishing Place 50 Hampshire St, Floor 5, Cambridge, Ma 02139 Usa
Reviewing status Peer reviewed
Institute(s) Institute of Pancreatic Islet Research (IPI)
POF-Topic(s) 90000 - German Center for Diabetes Research
Research field(s) Helmholtz Diabetes Center
PSP Element(s) G-502600-001
G-502600-002
G-502600-013
Grants German Center for Diabetes Research (DZD e.V.) - German Ministry for Education and Research
EFRE (European Fund for Regional Development)
Leona M. and Harry B. Helmsley Charitable Trust
JDRF International
Swiss State Secretariat for Education, Research, and Innovation
European Federation of Pharmacological Industries and Associations (EFPIA)
European Union's Framework Program Horizon 2020
Innovative Medicines Initiative 2 Joint Undertaking
German-Israeli Foundation for Scientific Research and Development (GIF)
Dresden International Graduate School for Biomedicine and Bioengineering (DIGS-BB)
DOI 10.1016/j.celrep.2024.113836
WOS ID 001193977800001
Scopus ID 85186764040
PubMed ID 38421874
Erfassungsdatum 2024-04-29
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