Identification of proteins undergoing tyrosine phosphorylation during mouse sperm capacitation.
Mammalian sperm are not able to fertilize immediately upon ejaculation; they become fertilization-competent after undergoing changes in the female reproductive tract collectively termed capacitation. Although it has been established that capacitation is associated with an increase in tyrosine phosphorylation, little is known about the role of this event in sperm function. In this work we used a combination of two dimensional gel electrophoresis and mass spectrometry to identify proteins that undergo tyrosine phosphorylation during capacitation. Some of the identified proteins are the mouse orthologues of human sperm proteins known to undergo tyrosine phosphorylation. Among them we identified VDAC, tubulin, PDH E1 beta chain, glutathione S-transferase, NADH dehydrogenase (ubiquinone) Fe-S protein 6, acrosin binding protein precursor (sp32), proteasome subunit alpha type 6b and cytochrome b-c1 complex. In addition to previously described proteins, we identified two testis-specific aldolases as substrates for tyrosine phosphorylation. Genomic and EST analyses suggest that these aldolases are retroposons expressed exclusively in the testis, as has been reported elsewhere. Because of the importance of glycolysis for sperm function, we hypothesize that tyrosine phosphorylation of these proteins can play a role in the regulation of glycolysis during capacitation. However, neither the Km nor the Vmax of aldolase changed as a function of capacitation when its enzymatic activity was assayed in vitro, suggesting other levels of regulation for aldolase function.
Arcelay
Enid
E
Department of Veterinary and Animal Sciences, University of Massachusetts, Amherst, MA, USA.
Salicioni
Ana M
AM
Wertheimer
Eva
E
Visconti
Pablo E
PE
D000818:Animals; D020224:Expressed Sequence Tags; D005634:Fructose-Bisphosphate Aldolase / Q000378:metabolism; D008297:Male; D051379:Mice; D008954:Models, Biological; D010766:Phosphorylation; D011506:Proteins / Q000737:chemistry*; D013075:Sperm Capacitation; D013094:Spermatozoa / Q000378:metabolism; D013737:Testis / Q000378:metabolism; D014404:Tubulin / Q000378:metabolism; D014443:Tyrosine / Q000737:chemistry*; D050994:Voltage-Dependent Anion Channels / Q000378:metabolism
D011506:Proteins; D014404:Tubulin; D050994:Voltage-Dependent Anion Channels; D014443:Tyrosine; D005634:Fructose-Bisphosphate Aldolase