Valosine containing protein is a substrate of cAMP-activated boar sperm tyrosine kinase.
Previously we reported that treatment of boar sperm with cAMP-elevating drugs induces tyrosine phosphorylation of a triton-insoluble 93 kDa protein (p93). We have isolated p93 by preparative SDS electrophoresis and blotting from urea-extracted boar sperm and identified it as a valosine containing protein (VCP) by mass spectrometry and microsequencing. With the use of antibodies to VCP and phosphotyrosine (pY) we found that both p93 and VCP are poorly extractable with triton and are solubilized in > 6 M urea. Furthermore, VCP and p93 overlap on one and two dimensional (1 and 2D) electrophoretic gels, supporting the identity of p93 as a tyrosine-phosphorylated population of VCP. According to immunofluorescence, VCP is localized along the entire sperm tail, in the posterior ring, distal equatorial segment, and postacrosome. In addition, 9-12% sperm contained VCP in the acrosome. The cAMP-elevating treatment did not alter VCP localization but induced tail tyrosine phosphorylation in 15% sperm cells. In those sperm, VCP and pY colocalized in connecting piece and posterior ring.
Geussova
Gizela
G
Department of Biology and Biochemistry of Fertilization, Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Videnska, Prague, Czech Republic.
Kalab
Petr
P
Peknicova
Jana
J
D000251:Adenosine Triphosphatases; D000595:Amino Acid Sequence; D000818:Animals; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D008297:Male; D008969:Molecular Sequence Data; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D013094:Spermatozoa / Q000201:enzymology*; D013552:Swine; D000074405:Valosin Containing Protein
D018797:Cell Cycle Proteins; D011505:Protein-Tyrosine Kinases; D000251:Adenosine Triphosphatases; D000074405:Valosin Containing Protein