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Proteomic analysis of insulin secretory granules in INS-1 cells by protein correlation profiling.

ABSTRACT:

Abstract

Insulin secretory granules (ISGs), a group of distinguishing organelles in pancreatic ? cells, are responsible for the storage and secretion of insulin to maintain blood glucose homeostasis. The molecular mechanisms of ISG biogenesis, maturation, transportation, and exocytosis are still largely unknown because the proteins involved in these distinct steps have not been fully identified. Subcellular fractionation by density gradient centrifugation has been successfully employed to analyze the proteomes of numerous organelles. However, use of this method to elucidate the ISG proteome is limited by co-fractionated contaminants because ISGs are very dynamic and have abundant exchanges or contacts with other organelles, such as the Golgi apparatus, lysosomes, and endosomes. In this study, we developed a new strategy for identifying ISG proteins by protein correlation profiling (PCP)-based proteomics, which included ISG purification by OptiPrep density gradient centrifugation, label-free quantitative proteome, and identification of ISG proteins by correlating fractionation profiles between candidates and known ISG markers. Using this approach, we were able to identify 81 ISG proteins. Among them, TM9SF3, a nine-transmembrane protein, was considered a high confidence ISG candidate protein highlighted in the PCP network. Further biochemical and immunofluorescence assays indicated that TM9SF3 localized in ISGs, suggesting that it is a potential new ISG marker.

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SUBMITTER: Li M

PROVIDER: S-EPMC6276070 | biostudies-literature | 2018

REPOSITORIES: biostudies-literature

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Proteomic analysis of insulin secretory granules in INS-1 cells by protein correlation profiling.

Li Min M Du Wen W Zhou Maoge M Zheng Li L Song Eli E Hou Junjie J

Biophysics reports 20180829 6

<h4>Abstract</h4>Insulin secretory granules (ISGs), a group of distinguishing organelles in pancreatic β cells, are responsible for the storage and secretion of insulin to maintain blood glucose homeostasis. The molecular mechanisms of ISG biogenesis, maturation, transportation, and exocytosis are still largely unknown because the proteins involved in these distinct steps have not been fully identified. Subcellular fractionation by density gradient centrifugation has been successfully employed t ...[more]

PMID: 30596141

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Dataset Information

Proteomic analysis of insulin secretory granules in INS-1 cells by protein correlation profiling.

Abstract

Graphical abstract

Publications

Proteomic analysis of insulin secretory granules in INS-1 cells by protein correlation profiling.

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