RT Journal Article
SR Electronic
T1 Purification of age-distinct insulin secretory granules through antigen restriction using the CLIP-tag
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.06.03.103770
DO 10.1101/2020.06.03.103770
A1 Neukam, Martin
A1 Ganß, Katharina
A1 Vasiljević, Jovana
A1 Broichhagen, Johannes
A1 Johnsson, Kai
A1 Kurth, Thomas
A1 Solimena, Michele
YR 2020
UL http://biorxiv.org/content/early/2020/06/03/2020.06.03.103770.abstract
AB Pancreatic islet beta cells employ secretory granules for the storage and glucose-stimulated release of the hormone insulin. The competence of an insulin granule for exocytosis depends on spatial and temporal variables such as its proximity to the plasma membrane as well as its age, with newly-generated granules being preferentially released. The molecular underpinnings for the control of these variables remain largely unknown and their uncovering is of high relevance for the study of diabetes, which results from deficient insulin secretion. However, we still lack a comprehensive view about the molecular composition of the insulin granules and how this may change over their lifetime. Here we report a strategy for the background-free purification of insulin secretory granules of distinct age from insulinoma INS-1 cells. We show that utilization of an immuno-based affinity approach for pulse-chase labeled insulin secretory granules, produces a highly enriched granular fraction. Our approach precludes typical contaminants from the solid phase and may be designed to purify secretory granules of a distinct age.Competing Interest StatementThe authors have declared no competing interest.