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

Neukam, M., Sala, P., Brunner, A.-D., Ganß, K., Palladini, A., Grzybek, M., et al. (2024). Purification of time-resolved insulin granules reveals proteomic and lipidomic changes during granule aging. Cell Reports, 43(3): 113836, pp. 1-23. doi:10.1016/j.celrep.2024.113836.

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Item Permalink: https://hdl.handle.net/21.11116/0000-000E-7E8D-7 Version Permalink: https://hdl.handle.net/21.11116/0000-000E-886F-D
Genre: Journal Article

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 Creators:
Neukam, Martin, Author
Sala, Pia, Author
Brunner, Andreas-David, Author
Ganß, Katharina, Author
Palladini, Alessandra, Author
Grzybek, Michal, Author
Topcheva, Oleksandra, Author
Vasiljević, Jovana, Author
Broichhagen, Johannes1, Author           
Johnsson, Kai1, Author           
Kurth, Thomas, Author
Mann, Matthias, Author
Coskun, Ünal, Author
Solimena, Michele, Author
Affiliations:
1Chemical Biology, Max Planck Institute for Medical Research, Max Planck Society, ou_2364732              

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Free keywords: ARHGAP1; CLIP tag; CP: Cell biology; CP: Metabolism; KIF5b; RAB3a; SNAP tag; granule aging; insulin granule; lipidomics; organelle purification; proteomics
 Abstract: 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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Language(s): eng - English
 Dates: 2023-12-292023-06-052024-02-052024-02-27
 Publication Status: Published online
 Pages: 23
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: URI: https://pubmed.ncbi.nlm.nih.gov/38421874/
DOI: 10.1016/j.celrep.2024.113836
 Degree: -

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Title: Cell Reports
Source Genre: Journal
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Publ. Info: Maryland Heights, MO : Cell Press
Pages: - Volume / Issue: 43 (3) Sequence Number: 113836 Start / End Page: 1 - 23 Identifier: ISSN: 2211-1247
CoNE: https://pure.mpg.de/cone/journals/resource/2211-1247