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Identification of increased amounts of eppin protein complex components in sperm cells of diabetic and obese individuals by difference gel electrophoresis

Item Type:Article
Title:Identification of increased amounts of eppin protein complex components in sperm cells of diabetic and obese individuals by difference gel electrophoresis
Creators Name:Paasch, U. and Heidenreich, F. and Pursche, T. and Kuhlisch, E. and Kettner, K. and Grunewald, S. and Kratzsch, J. and Dittmar, G. and Glander, H.J. and Hoflack, B. and Kriegel, T.M.
Abstract:Metabolic disorders like diabetes mellitus and obesity may compromise the fertility of men and women. To unveil disease-associated proteomic changes potentially affecting male fertility, the proteomes of sperm cells from type-1-diabetic, type-2-diabetic, non-diabetic obese and clinically healthy individuals were comparatively analyzed by differ-ence gel electrophoresis (DIGE). The adaptation of a general protein extraction proce-dure to the solubilization of proteins from sperm cells allowed for the resolution of 3,187 fluorescent spots in the DIGE image of the master gel, which contained the entirety of solubilized sperm proteins. Comparison of the pathological and reference proteomes by applying an average abundance ratio setting of 1.6 and a p≤0.05 criterion resulted in the identification of 79 fluorescent spots containing proteins that were present at significantly changed levels in the sperm cells. Biometric evaluation of the fluorescence data followed by mass spectrometric protein identification revealed altered levels of 12, 71 and 13 pro-tein species in the proteomes of the type-1-diabetic, type-2-diabetic and non-diabetic obese patients, respectively, with considerably enhanced levels of the same set of one molecular form of semenogelin-1, one form of clusterin and two forms of lactotransferrin in each group of pathological samples. Remarkably,galactosidase-1-like protein was the only protein that was detected at decreased levels in all three pathological situations. The former three proteins are part of the eppin (epididymal proteinase inhibitor) protein complex (EPC), which is thought to fulfill fertilization-related functions, such as ejaculate sperm protection, motility regulation and gain of competence for acrosome reaction, while the putative role of the latter protein to function as a glycosyl hydrolase during sperm maturation remains to be explored at the protein/enzyme level. The strikingly similar differences detected in the three groups of pathological sperm proteomes reflect a disease-associated enhanced formation of predominantly proteolytically modified forms of the EPC components, possibly as a response to enduring hyperglycemia and enhanced oxidative stress.
Keywords:DIGE, Diabetes, EPC, Eppin Protein Complex, Human, Obesity, Proteome, Sperm
Source:Molecular & Cellular Proteomics
Publisher:American Society for Biochemistry and Molecular Biology (U.S.A.)
Page Range:M110.007187
Date:August 2011
Official Publication:https://doi.org/10.1074/mcp.M110.007187
PubMed:View item in PubMed

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