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Identification of post‐translational modifications that occur during sperm maturation using difference in two‐dimensional gel electrophoresis
Author(s) -
Baker Mark A.,
Witherdin Roxanne,
Hetherington Louise,
CunninghamSmith Kelly,
Aitken R. John
Publication year - 2005
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200401100
Subject(s) - epididymis , difference gel electrophoresis , sperm , gel electrophoresis , phosphorylation , serine , motility , biology , polyacrylamide gel electrophoresis , proteomics , proteome , protein subunit , sperm motility , microbiology and biotechnology , biochemistry , chemistry , genetics , enzyme , gene
Difference in two‐dimensional (2‐D) gel electrophoresis (DIGE) is a novel method for analyzing up to three samples in one 2‐D gel and using the information gained to study post‐translational modifications of proteins. We describe the use of DIGE to isolate and characterize those proteins that undergo processing in spermatozoa as they transit the epididymal tract. We find up to 60 protein spots are significantly modified as sperm traverse the epididymis. In this article, we report eight unambiguous protein identifications and demonstrate that one protein, the β‐subunit of the mitochondrial F1‐ATPase, is serine‐phosphorylated as sperm undergo epididymal maturation. We suggest that phosphorylation of this particular protein in a cAMP‐dependent manner may contribute to the mechanisms by which motility is conferred upon spermatozoa.