Phosphorylation of SNAP-25 on Serine-187 is Induced by Secretagogues in Insulin-secreting Cells, but is Not Correlated with Insulin Secretion

Overview

Journal Biochem J

Specialty Biochemistry

Date 2002 Aug 8

PMID 12164783

Citations 10

Authors

Carmen Gonelle-Gispert

Maria Costa

Masami Takahashi

Karin Sadoul

Philippe Halban

Affiliations

Soon will be listed here.

Abstract

The tSNARE (the target-membrane soluble NSF-attachment protein receptor, where NSF is N -ethylmaleimide-sensitive fusion protein) synaptosomal-associated protein of 25 kDa (SNAP-25) is implicated in regulated insulin secretion. In pheochromocytoma PC12 cells, SNAP-25 is phosphorylated at Ser(187), 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 Ser(187) 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 Ser(187) 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, Ser(187) 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 Ser(187)-->Ala or Ser(187)-->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 Ser(187) 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 Ser(187) occurs in insulin-secreting cells and is mediated by PKC, it does not appear to play a major role in regulated insulin secretion.

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References

Malaisse W, Dunlop M, Mathias P, Malaisse-Lagae F, Sener A . Stimulation of protein kinase C and insulin release by 1-oleoyl-2-acetyl-glycerol. Eur J Biochem. 1985; 149(1):23-7. DOI: 10.1111/j.1432-1033.1985.tb08887.x. View

Foster L, Yeung B, Mohtashami M, Ross K, Trimble W, Klip A . Binary interactions of the SNARE proteins syntaxin-4, SNAP23, and VAMP-2 and their regulation by phosphorylation. Biochemistry. 1998; 37(31):11089-96. DOI: 10.1021/bi980253t. View

Vitale M, Seward E, TRIFARO J . Chromaffin cell cortical actin network dynamics control the size of the release-ready vesicle pool and the initial rate of exocytosis. Neuron. 1995; 14(2):353-63. DOI: 10.1016/0896-6273(95)90291-0. View

Shimazaki Y, Nishiki T, Omori A, Sekiguchi M, Kamata Y, Kozaki S . Phosphorylation of 25-kDa synaptosome-associated protein. Possible involvement in protein kinase C-mediated regulation of neurotransmitter release. J Biol Chem. 1996; 271(24):14548-53. DOI: 10.1074/jbc.271.24.14548. View

Laemmli U . Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970; 227(5259):680-5. DOI: 10.1038/227680a0. View

Asfari M, Janjic D, Meda P, Li G, Halban P, Wollheim C . Establishment of 2-mercaptoethanol-dependent differentiated insulin-secreting cell lines. Endocrinology. 1992; 130(1):167-78. DOI: 10.1210/endo.130.1.1370150. View

Lin R, Scheller R . Structural organization of the synaptic exocytosis core complex. Neuron. 1997; 19(5):1087-94. DOI: 10.1016/s0896-6273(00)80399-2. View

Nielander H, Onofri F, Valtorta F, Schiavo G, Montecucco C, Greengard P . Phosphorylation of VAMP/synaptobrevin in synaptic vesicles by endogenous protein kinases. J Neurochem. 1995; 65(4):1712-20. DOI: 10.1046/j.1471-4159.1995.65041712.x. View

Ganesan S, Calle R, Zawalich K, Smallwood J, Zawalich W, Rasmussen H . Glucose-induced translocation of protein kinase C in rat pancreatic islets. Proc Natl Acad Sci U S A. 1990; 87(24):9893-7. PMC: 55280. DOI: 10.1073/pnas.87.24.9893. View

10.

Sollner T, Whiteheart S, Brunner M, Erdjument-Bromage H, Geromanos S, Tempst P . SNAP receptors implicated in vesicle targeting and fusion. Nature. 1993; 362(6418):318-24. DOI: 10.1038/362318a0. View

11.

Sadoul K, LANG J, Montecucco C, Weller U, Regazzi R, Catsicas S . SNAP-25 is expressed in islets of Langerhans and is involved in insulin release. J Cell Biol. 1995; 128(6):1019-28. PMC: 2120411. DOI: 10.1083/jcb.128.6.1019. View

12.

Braiman L, Alt A, Kuroki T, Ohba M, Bak A, Tennenbaum T . Activation of protein kinase C zeta induces serine phosphorylation of VAMP2 in the GLUT4 compartment and increases glucose transport in skeletal muscle. Mol Cell Biol. 2001; 21(22):7852-61. PMC: 99955. DOI: 10.1128/MCB.21.22.7852-7861.2001. View

13.

Wollheim C, Dunne M, Bruzzone R, Pozzan T, Petersen O . Activators of protein kinase C depolarize insulin-secreting cells by closing K+ channels. EMBO J. 1988; 7(8):2443-9. PMC: 457113. DOI: 10.1002/j.1460-2075.1988.tb03090.x. View

14.

Jones P, Persaud S . Protein kinases, protein phosphorylation, and the regulation of insulin secretion from pancreatic beta-cells. Endocr Rev. 1998; 19(4):429-61. DOI: 10.1210/edrv.19.4.0339. View

15.

RISINGER C, Bennett M . Differential phosphorylation of syntaxin and synaptosome-associated protein of 25 kDa (SNAP-25) isoforms. J Neurochem. 1999; 72(2):614-24. DOI: 10.1046/j.1471-4159.1999.0720614.x. View

16.

Kataoka M, Kuwahara R, Iwasaki S, Takahashi M . Nerve growth factor-induced phosphorylation of SNAP-25 in PC12 cells: a possible involvement in the regulation of SNAP-25 localization. J Neurochem. 2000; 74(5):2058-66. DOI: 10.1046/j.1471-4159.2000.0742058.x. View

17.

Pang D, Wang J, Valtorta F, Benfenati F, Greengard P . Protein tyrosine phosphorylation in synaptic vesicles. Proc Natl Acad Sci U S A. 1988; 85(3):762-6. PMC: 279635. DOI: 10.1073/pnas.85.3.762. View

18.

De Camilli P, Benfenati F, Valtorta F, Greengard P . The synapsins. Annu Rev Cell Biol. 1990; 6:433-60. DOI: 10.1146/annurev.cb.06.110190.002245. View

19.

Cabaniols J, Ravichandran V, Roche P . Phosphorylation of SNAP-23 by the novel kinase SNAK regulates t-SNARE complex assembly. Mol Biol Cell. 1999; 10(12):4033-41. PMC: 25741. DOI: 10.1091/mbc.10.12.4033. View

20.

Halban P, Niemann H, Palmer M, Catsicas S, Sadoul K . SNAP-25a and -25b isoforms are both expressed in insulin-secreting cells and can function in insulin secretion. Biochem J. 1999; 339 ( Pt 1):159-65. PMC: 1220140. View