Two-dimensional gel electrophoresis of CyDye-labeled human sperm proteomes (DIGE) and identification of diabetes- and obesity-associated sperm proteins.

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Two-dimensional gel electrophoresis of CyDye-labeled human sperm proteomes (DIGE) and identification of diabetes- and obesity-associated sperm proteins. Two-dimensional gel electrophoresis of CyDye-labeled human sperm proteomes (DIGE) and identification of diabetes- and obesity-associated sperm proteins. Fluorescent spots containing proteins that occur at pathologically altered concentrations are labeled and annotated in red (type-1 diabetes), blue (type-2 diabetes), and yellow (obesity). Proteins that show very similar abundance changes in all three metabolic disorders are labeled and annotated in white, whereas proteins showing very similar abundance changes in two metabolic disorders are labeled and annotated in green. A, DIGE image of the internal standard comprising all reference and sample proteins (master gel). B–D: Illustration of proteomic changes for type-1 diabetic (B), type-2 diabetic (C) and non-diabetic obese (D) patients in the master gel image. Boundaries surround fluorescent spots exhibiting significantly increased (↑) or decreased (↓) protein volumes or, in case of low-abundance proteins, their respective position in the two-dimensional gel. The dot within each boundary is marking the center of protein mass, which generally represents the optimal picking location for protein identification. In Fig. 1C, the proteins listed in Table III are labeled, whereas the complete set of proteins associated with type-2 diabetes is available in the “Supplemental data” section. Identified proteins are annotated by their gene name according to the Swiss-Prot protein database and by their spot ID used in Tables II–IV and Table SI where full protein names are also given. Uwe Paasch et al. Mol Cell Proteomics 2011;10:M110.007187 © 2011 by The American Society for Biochemistry and Molecular Biology, Inc.