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Proteomics analysis of insulin secretory granules

Fukuda, Mitsonuri
Published in Molecular & cellular proteomics. 2007, vol. 6, no. 6, p. 1007-17
Abstract Insulin secretory granules (ISGs) are cytoplasmic organelles of pancreatic beta-cells. They are responsible for the storage and secretion of insulin. To date, only about 30 different proteins have been clearly described to be associated with these organelles. However, data from two-dimensional gel electrophoresis analyses suggested that almost 150 different polypeptides might be present within ISGs. The elucidation of the identity and function of the ISG proteins by proteomics strategies would be of considerable help to further understand some of the underlying mechanisms implicated in ISG biogenesis and trafficking. Furthermore it should give the bases to the comprehension of impaired insulin secretion observed during diabetes. A proteomics analysis of an enriched insulin granule fraction from the rat insulin-secreting cell line INS-1E was performed. The efficacy of the fractionation procedure was assessed by Western blot and electron microscopy. Proteins of the ISG fraction were separated by SDS-PAGE, excised from consecutive gel slices, and tryptically digested. Peptides were analyzed by nano-LC-ESI-MS/MS. This strategy identified 130 different proteins that were classified into four structural groups including intravesicular proteins, membrane proteins, novel proteins, and other proteins. Confocal microscopy analysis demonstrated the association of Rab37 and VAMP8 with ISGs in INS-1E cells. In conclusion, the present study identified 130 proteins from which 110 are new proteins associated with ISGs. The elucidation of their role will further help in the understanding of the mechanisms governing impaired insulin secretion during diabetes.
Keywords AnimalsImmunoblottingInsulin/secretionMass SpectrometryMiceProtein TransportProteins/analysis/chemistry/isolation & purification/secretionProteomicsRatsReproducibility of ResultsSecretory Vesicles/chemistry/ultrastructureSubcellular Fractions/chemistry
PMID: 17317658
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Article (Published version) (411 Kb) - public document Free access
Article (Published version) (1 MB) - public document Free access
Research groups Chimie et protéomique clinique (270)
Groupe de Protéomique biomédicale (635)
Obésité et syndrome métabolique (803)
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BRUNNER, Yannick et al. Proteomics analysis of insulin secretory granules. In: Molecular and Cellular Proteomics, 2007, vol. 6, n° 6, p. 1007-17. doi: 10.1074/mcp.M600443-MCP200 https://archive-ouverte.unige.ch/unige:33456

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Deposited on : 2014-01-20

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