Novel tyrosine-phosphorylated post-pyruvate metabolic enzyme, dihydrolipoamide dehydrogenase, involved in capacitation of hamster spermatozoa.
Capacitation is a process that confers fertilizing ability to spermatozoa and this critical event occurs in the development of mammalian spermatozoa during their transit through the female reproductive tract and precedes fertilization. Because spermatozoa are relatively silent in transcription and translation, posttranslational modifications perform the regulatory functions in these cells during capacitation. In this report, we identify a candidate protein, dihydrolipoamide dehydrogenase, which is a post-pyruvate metabolic enzyme, exhibiting tyrosine phosphorylation during hamster spermatozoal capacitation. This is the first report showing dihydrolipoamide dehydrogenase as a phosphoprotein. The cDNA sequence of hamster testes dihydrolipoamide dehydrogenase does not show any variation from the already reported mammalian dihydrolipoamide dehydrogenases. Downregulation of the activity of the hamster spermatozoal enzyme by its specific inhibitor, 5-methoxyindole-2-carboxylic acid, blocks acrosome reaction completely and hyperactivation partially, confirming the role of dihydrolipoamide dehydrogenase in hamster spermatozoal capacitation. We also delineate the temporal involvement of glucose and pyruvate-lactate, showing that the former is required in the earlier stages and the latter for the later stages of hamster spermatozoal capacitation. The essentiality of pyruvate-lactate during hyperactivation and acrosome reaction necessitates the involvement of the post-pyruvate-lactate enzyme, dihydrolipoamide dehydrogenase.
Mitra
Kasturi
K
Centre for Cellular and Molecular Biology, Hyderabad 500 007, India.
Shivaji
S
S
D020101:Acrosome Reaction / Q000187:drug effects / Q000502:physiology; D000595:Amino Acid Sequence; D000818:Animals; D002118:Calcium / Q000494:pharmacology; D006224:Cricetinae; D008058:Dihydrolipoamide Dehydrogenase / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005947:Glucose / Q000378:metabolism; D007211:Indoles / Q000494:pharmacology; D019344:Lactic Acid / Q000378:metabolism; D008297:Male; D008647:Mesocricetus; D008969:Molecular Sequence Data; D010766:Phosphorylation; D019289:Pyruvic Acid / Q000378:metabolism*; D013075:Sperm Capacitation / Q000502:physiology*; D013094:Spermatozoa / Q000378:metabolism / Q000502:physiology*; D013997:Time Factors; D014443:Tyrosine / Q000378:metabolism*
D004791:Enzyme Inhibitors; D007211:Indoles; C008400:5-methoxyindole-2-carboxylic acid; D019344:Lactic Acid; D014443:Tyrosine; D019289:Pyruvic Acid; D008058:Dihydrolipoamide Dehydrogenase; D005947:Glucose; D002118:Calcium