Hexokinase present in human sperm is not tyrosine phosphorylated but its antibodies affect fertilizing capacity.
The present study was conducted to investigate the presence of hexokinase in human sperm cell, its subcellular localization, its modulation and role in capacitation/acrosome reaction, and tyrosine kinase activity, if any. These studies were conducted using antibodies (Ab) raised against rat brain hexokinase (type I isozyme) that have significant cross-reaction with various human tissue hexokinases and neutralize the catalytic activity of the enzyme. Hexokinase Ab reacted with acrosomal, mid-piece and tail regions of methanol-fixed (approximately 70-80%) and live (approximately 35-52%) sperm in the indirect immunofluorescence technique (IFT) and the immunobead binding technique (IBT), respectively. Hexokinase Ab specifically recognized a band of 116 kDa on the Western blot of detergent-solubilized human sperm preparation that was different from the 95-kDa phosphotyrosine protein. Hexokinase Ab caused a significant (P < 0.01 to < 0.001) and concentration-dependent inhibition of human sperm penetration of zona-free hamster oocytes in the sperm penetration assay (SPA). These data indicate that the hexokinase of 116-kDa molecular weight is present in acrosomal, mid-piece, and tail regions of human sperm. The sperm hexokinase is a glycoprotein that is different from the 95-kDa phosphotyrosine protein and is not phosphorylated at tyrosine residues; however, its antibodies cause agglutination and a concentration-dependent inhibition of fertilizing capacity of human sperm.
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Department of Obstetrics and Gynecology, Albert Einstein College of Medicine, Bronx, New York 10461, USA.
Morte
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C
Ahmad
K
K
Martinez
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P
D000906:Antibodies / Q000032:analysis; D015153:Blotting, Western; D003429:Cross Reactions; D005260:Female; D005306:Fertilization / Q000502:physiology*; D005455:Fluorescent Antibody Technique; D006593:Hexokinase / Q000032:analysis* / Q000276:immunology; D006801:Humans; D008297:Male; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000737:chemistry; D013075:Sperm Capacitation / Q000502:physiology; D013084:Sperm-Ovum Interactions / Q000502:physiology; D013094:Spermatozoa / Q000201:enzymology* / Q000502:physiology / Q000648:ultrastructure
D000906:Antibodies; D006593:Hexokinase; D011505:Protein-Tyrosine Kinases