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

ABSTRACT: Endocrine cells employ regulated exocytosis of secretory granules to secrete hormones and neurotransmitters. Secretory granule exocytosis depends on spatio-temporal 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 about 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

INSTRUMENT(S):

ORGANISM(S): Rattus Norvegicus (rat)

TISSUE(S): Pancreas

DISEASE(S): Diabetes Mellitus

SUBMITTER: Matthias Mann  

LAB HEAD: Matthias Mann

PROVIDER: PXD050087 | Pride | 2024-04-15

REPOSITORIES: Pride