Click
here to close Hello! We notice that
you are using Internet Explorer, which is not supported by Echinobase
and may cause the site to display incorrectly. We suggest using a
current version of Chrome,
FireFox,
or Safari.
???displayArticle.abstract???
Calmodulin (CaM) affinity chromatography of a detergent extract of sea urchin sperm yielded approximately 20 major proteins. One of these proteins, of Mr 190,000, was purified and used to immunize rabbits. After absorption with living sperm, the serum reacted monospecifically on one- and two-dimensional Western immunoblots with the Mr 190,000 protein. The anti-190-kD serum inhibited 94% of the adenylate cyclase (AC) activity of the CaM eluate. An immunoaffinity column removed 95% of the AC activity, and the purified (but inactive) Mr 190,000 protein was eluted from the column. The antiserum also inhibited 23% of the activity of bovine brain CaM-sensitive AC and 90% of the activity of horse sperm CaM-sensitive AC. These data support the hypothesis that the Mr 190,000 protein is sea urchin sperm AC. Although this AC bound to CaM, it was not possible to demonstrate directly a Ca2+ or CaM sensitivity. However, two CaM antagonists, calmidazolium and chlorpromazine, both inhibited AC activity, and the inhibition was released by added CaM, suggesting the possibility of regulation of this AC by CaM. Indirect immunofluorescence showed the Mr 190,000 protein to be highly concentrated on only the proximal half of the sea urchin sperm flagellum. This asymmetric localization of AC may be important to its function in flagellar motility. This is the first report of the identification of an AC from animal spermatozoa.
Brokaw,
Regulation of sperm flagellar motility by calcium and cAMP-dependent phosphorylation.
1987, Pubmed,
Echinobase
Brokaw,
Regulation of sperm flagellar motility by calcium and cAMP-dependent phosphorylation.
1987,
Pubmed
,
Echinobase
Garbers,
The regulation of spermatozoa by calcium cyclic nucleotides.
1980,
Pubmed
Gross,
Calmodulin-mediated adenylate cyclase from mammalian sperm.
1987,
Pubmed
Kopf,
Characterization of a calmodulin-stimulated adenylate cyclase from abalone spermatozoa.
1984,
Pubmed
Kopf,
Characterization of a calcium-modulated adenylate cyclase from abalone spermatozoa.
1985,
Pubmed
LOWRY,
Protein measurement with the Folin phenol reagent.
1951,
Pubmed
Laemmli,
Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
1970,
Pubmed
Lindemann,
Regulation of mammalian sperm motility.
1989,
Pubmed
Macfarlane,
Two dimensional benzyldimethyl-n-hexadecylammonium chloride----sodium dodecyl sulfate preparative polyacrylamide gel electrophoresis: a high capacity high resolution technique for the purification of proteins from complex mixtures.
1989,
Pubmed
Morrissey,
Silver stain for proteins in polyacrylamide gels: a modified procedure with enhanced uniform sensitivity.
1981,
Pubmed
Newman,
Lymphoid receptors for transferrin.
1987,
Pubmed
Nishioka,
Changing localizations of site-specific surface antigens during sea urchin spermiogenesis.
1987,
Pubmed
,
Echinobase
O'Neil,
How calmodulin binds its targets: sequence independent recognition of amphiphilic alpha-helices.
1990,
Pubmed
Pfeuffer,
Olfactory adenylyl cyclase. Identification and purification of a novel enzyme form.
1989,
Pubmed
Podell,
Isolation and characterization of a plasma membrane fraction from sea urchin sperm exhibiting species specific recognition of the egg surface.
1984,
Pubmed
,
Echinobase
Sano,
Subcellular localizations of guanylate cyclase and 3',5'-cyclic nucleotide phosphodiesterase in sea urchin sperm.
1976,
Pubmed
,
Echinobase
Smith,
Measurement of protein using bicinchoninic acid.
1985,
Pubmed
Tash,
Protein phosphorylation: the second messenger signal transducer of flagellar motility.
1989,
Pubmed
Tash,
Ca2+ regulation of sperm axonemal motility.
1987,
Pubmed
Tombes,
Sea urchin sperm creatine kinase: the flagellar isozyme is a microtubule-associated protein.
1988,
Pubmed
,
Echinobase
Toowicharanont,
Regional differentiation of the sea urchin sperm plasma membrane.
1988,
Pubmed
,
Echinobase
Trimmer,
Monoclonal antibodies increase intracellular Ca2+ in sea urchin spermatozoa.
1986,
Pubmed
,
Echinobase
Vacquier,
Stoichiometry of phosphate loss from sea urchin sperm guanylate cyclase during fertilization.
1986,
Pubmed
,
Echinobase
Vacquier,
Monoclonal antibodies to a membrane glycoprotein induce the phosphorylation of histone H1 in sea urchin spermatozoa.
1988,
Pubmed
,
Echinobase
Ward,
Phosphorylation of membrane-bound guanylate cyclase of sea urchin spermatozoa.
1986,
Pubmed
,
Echinobase
Yamawaki-Kataoka,
Adenylate cyclases in yeast: a comparison of the genes from Schizosaccharomyces pombe and Saccharomyces cerevisiae.
1989,
Pubmed
Yeager,
Purification of the calmodulin-sensitive adenylate cyclase from bovine cerebral cortex.
1985,
Pubmed
