Protein kinase A activity and protein phosphorylation during the mouse sperm acrosomal reaction.
Using two-dimensional gel electrophoresis, changes in protein phosphorylation caused by cyclic nucleotide-dependent protein kinases were analyzed with or without exposure to a protein kinase inhibitor, H-8, during the mouse sperm acrosomal reaction. The acrosomal reaction, induced by the treatment of sperm with dibutyryl cyclic AMP or dibutyryl cyclic GMP, was inhibited by H-8. The activities of cyclic AMP-dependent protein kinase (PKA) and cyclic GMP-dependent protein kinase induced by the sperm extract were also inhibited by H-8. When endogenous PKA in sperm was activated by the addition of cyclic AMP, a 45-kDa protein spot identified by electrophoresis indicated the occurrence of phosphorylation in vivo. Furthermore, the enhanced phosphorylation of the 45-kDa protein spot was inhibited by H-8. These results suggest that the PKA-catalyzed phosphorylation of the 45-kDa protein may be involved in the regulation of the mouse sperm acrosomal reaction.
Kuji
N
N
Department of Obstetrics and Gynecology, Keio University, School of Medicine, Tokyo, Japan. naoaki@sc.itc.keio.ac.jp
Tanaka
Y
Y
Komatsu
S
S
Yoshimura
Y
Y
D020101:Acrosome Reaction; D000818:Animals; D003994:Bucladesine / Q000494:pharmacology; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism*; D003995:Dibutyryl Cyclic GMP / Q000494:pharmacology; D015180:Electrophoresis, Gel, Two-Dimensional; D008297:Male; D051379:Mice; D010766:Phosphorylation; D011506:Proteins / Q000378:metabolism*; D013094:Spermatozoa / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism / Q000502:physiology*
D011506:Proteins; D003995:Dibutyryl Cyclic GMP; D003994:Bucladesine; D017868:Cyclic AMP-Dependent Protein Kinases