Redox control of changes in protein sulfhydryl levels during human sperm capacitation.
Capacitation, the series of transformations that spermatozoa undergo to become fertile, is regulated by reactive oxygen species (ROS) and associated with an increase in the sulfhydryl content of Triton-soluble proteins. Our aims were to determine the fate of sulfhydryl groups in Triton-soluble proteins from capacitating human spermatozoa using two-dimensional (2D) gel electrophoresis, to evaluate the role of ROS in the changes observed, and to correlate the time course of the changes with that of the sperm generation of O(2)(*)(-). Triton-soluble proteins of control and capacitating human spermatozoa were labeled with 3-(N-maleimidylpropionyl) biocytin, separated by 2D gel electrophoresis, and probed with horseradish peroxidase-conjugated streptavidin. The sulfhydryl content of 10 out of the 14 proteins studied (pI: 4-7) was modified by the induction of capacitation, and the increases (by 200-400%, five proteins) and decreases (by 45-95%, five proteins) were prevented by superoxide dismutase and/or catalase. The alterations in protein sulfhydryl content occurred within 5-15 min but were reversed within 30-120 min. Three capacitation inducers triggered similar modifications. Therefore, human sperm capacitation is associated with rapid and reversible changes in protein sulfhydryl groups that appear to be redox regulated. The number of proteins affected, the types, and the kinetics of changes emphasize the complexity of sperm capacitation.
de Lamirande
Eve
E
Urology Research Laboratory, Royal Victoria Hospital, Montréal, Québec, Canada. edelamirande@hotmail.com
Gagnon
Claude
C
D002374:Catalase / Q000494:pharmacology; D015180:Electrophoresis, Gel, Two-Dimensional; D005312:Fetal Blood / Q000378:metabolism; D006801:Humans; D008239:Lysine / Q000031:analogs & derivatives*; D008297:Male; D008301:Maleimides; D010084:Oxidation-Reduction; D029607:Seminal Vesicle Secretory Proteins / Q000737:chemistry / Q000378:metabolism*; D013075:Sperm Capacitation; D013094:Spermatozoa / Q000378:metabolism*; D013438:Sulfhydryl Compounds / Q000378:metabolism*; D013439:Sulfhydryl Reagents / Q000494:pharmacology; D013482:Superoxide Dismutase / Q000494:pharmacology; D013481:Superoxides / Q000378:metabolism*
D008301:Maleimides; D029607:Seminal Vesicle Secretory Proteins; D013438:Sulfhydryl Compounds; D013439:Sulfhydryl Reagents; D013481:Superoxides; C047272:3-(N-maleimidopropionyl)biocytin; D002374:Catalase; D013482:Superoxide Dismutase; D008239:Lysine