Improved characterization of the insulin secretory granule proteomes

Biomedical Proteomics Research Group, Department of Human Protein Sciences, University Medical Center, Geneva, Switzerland.
Journal of proteomics (Impact Factor: 3.89). 04/2012; 75(15):4620-31. DOI: 10.1016/j.jprot.2012.04.023
Source: PubMed


Insulin secretory granules (ISGs) are pivotal organelles of pancreatic ß-cells and represent a key participant to glucose homeostasis. Indeed, insulin is packed and processed within these vesicles before its release by exocytosis. It is therefore crucial to acquire qualitative and quantitative data on the ISG proteome, in order to increase our knowledge on ISG biogenesis, maturation and exocytosis. Despites efforts made in the past years, the coverage of the ISG proteome is still incomplete and comprises many potential protein contaminants most likely coming from suboptimal sample preparations. We developed here a 3-step gradient purification procedure combined to Stable Isotope Labeling with Amino acids in Cell culture (SILAC) to further characterize the ISG protein content. Our results allowed to build three complementary proteomes containing 1/ proteins which are enriched in mature ISGs, 2/ proteins sharing multiple localizations including ISGs, and finally 3/ proteins sorted out from immature ISGs and/or co-purifying contaminants. As a proof of concept, the ProSAAS, a neuronal protein found in ISGs was further characterized and its granular localization proved. ProSAAS might represent a novel potential target allowing to better understand the defaults in insulin processing and secretion observed during type 2 diabetes progression. This article is part of a special issue entitled: Translational Proteomics.

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Available from: Domitille Schvartz, Dec 18, 2013
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    • "In the first work, they only enriched total ISGs, without further distinguishing between iISGs and mISGs (Brunner et al. 2007). In the second study, they used a three-step gradient purification procedure to obtain both iISGs and mISGs; however, the properties and purity of the final fractions were not well characterized (Schvartz et al. 2012). Here, we developed a simpler enrichment method and achieved better separation, as verified by many specific markers. "
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