The tSNARE (the target-membrane soluble NSF-attachment protein receptor, where NSF is N-ethylmaleimide-sensitive fusion protein) synaptosomal-associated protein of 25kDa (SNAP-25) is implicated in regulated insulin secretion. In pheochromocytoma PC12 cells, SNAP-25 is phosphorylated at Ser187, which lies in a region that is important for its function. The aims of the present study were to determine whether SNAP-25 is phosphorylated at Ser187 in insulin-secreting cells and, if so, whether this is important for regulated insulin secretion. The major findings are: (i) SNAP-25 is rapidly and reversibly phosphorylated on Ser187 in both rat insulinoma INS-1 cells and rat islets in response to the phorbol ester, PMA; (ii) less than 35% of SNAP-25 in INS-1 cells is phosphorylated in response to PMA, and phosphorylation is limited to plasma-membrane-associated SNAP-25; (iii) both SNAP-25 isoforms (a and b) are phosphorylated, with 1.8-fold greater phosphorylation for SNAP-25b in response to PMA; (iv) in rat islets, Ser187 phosphorylation is stimulated by glucose or carbachol, albeit to a lesser extent than by PMA, but not by cAMP; (v) insulin secretion from botulinum neurotoxin E-treated hamster insulinoma tumour (HIT) cells, transfected with toxin-resistant Ser187→Ala or Ser187→Asp mutant SNAP-25, was similar to that of wild-type HIT cells. Furthermore, in rat islets no correlation was found between the extent of SNAP-25 phosphorylation at Ser187 in response to secretagogues and stimulation of insulin release; (vi) use of protein kinase C (PKC) inhibitors suggests that glucose stimulates SNAP-25 phosphorylation via conventional and non-conventional PKC isoforms. In summary, although SNAP-25 phosphorylation at Ser187 occurs in insulin-secreting cells and is mediated by PKC, it does not appear to play a major role in regulated insulin secretion.
Skip Nav Destination
Article navigation
November 2002
- PDF Icon PDF LinkFront Matter
Research Article|
November 15 2002
Phosphorylation of SNAP-25 on serine-187 is induced by secretagogues in insulin-secreting cells, but is not correlated with insulin secretion
Carmen GONELLE-GISPERT;
Carmen GONELLE-GISPERT
1
∗Laboratoires de Recherche Louis Jeantet, Centre Médical Universitaire, rue Michel Servet, CH-1211 Geneva 4, Switzerland
1To whom correspondence should be addressed (e-mail carmen.gonelle@medecine.unige.ch).
Search for other works by this author on:
Maria COSTA;
Maria COSTA
∗Laboratoires de Recherche Louis Jeantet, Centre Médical Universitaire, rue Michel Servet, CH-1211 Geneva 4, Switzerland
Search for other works by this author on:
Masami TAKAHASHI;
Masami TAKAHASHI
†Project Research Center, Mitsubishikasei Institute of Life Sciences, Tokyo 194-8511, Japan,
Search for other works by this author on:
Karin SADOUL;
Karin SADOUL
‡LEDAC, Institut Albert Bonniot, Faculté de Médecine, Domaine de la Merci, 38706 La Tronche Cedex, France
Search for other works by this author on:
Philippe HALBAN
Philippe HALBAN
∗Laboratoires de Recherche Louis Jeantet, Centre Médical Universitaire, rue Michel Servet, CH-1211 Geneva 4, Switzerland
Search for other works by this author on:
Publisher: Portland Press Ltd
Received:
June 11 2002
Revision Received:
July 29 2002
Accepted:
August 06 2002
Accepted Manuscript online:
August 06 2002
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London ©2002
2002
Biochem J (2002) 368 (1): 223–232.
Article history
Received:
June 11 2002
Revision Received:
July 29 2002
Accepted:
August 06 2002
Accepted Manuscript online:
August 06 2002
Citation
Carmen GONELLE-GISPERT, Maria COSTA, Masami TAKAHASHI, Karin SADOUL, Philippe HALBAN; Phosphorylation of SNAP-25 on serine-187 is induced by secretagogues in insulin-secreting cells, but is not correlated with insulin secretion. Biochem J 15 November 2002; 368 (1): 223–232. doi: https://doi.org/10.1042/bj20020896
Download citation file:
Sign in
Don't already have an account? Register
Sign in to your personal account
You could not be signed in. Please check your email address / username and password and try again.
Captcha Validation Error. Please try again.