Expression of Sperm-Activating Peptide IV Receptor-Associated Membrane Guanylyl Cyclase in the Testis of the Sea Urchin Diadema setosum

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  • Expression of Sperm-Activating Peptide IV Receptor-Associated Membrane Guanylyl Cyclase in the Testis of the Sea Urchin Diadema setosum(Biochemistry) :

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We obtained the full-length cDNA clone (DsPTGC04) encoding a membrane guanylyl cyclase expressed in the testis of the sea urchin Diadema setosum, the egg jelly of which contains sperm-activating peptide IV. The cDNA was 4305 bp long and an open reading frame predicted a protein of 1127 amino acids including an apparent signal peptide of 24 residues. The mature protein of 1103 amino acids is composed of a single transmembrane domain of 25 amino acids that divides the mature protein (Mw 123818) into an amino-terminal, extracellular domain of 484 amino acids and a carboxyl-terminal, intracellular domain of 594 amino acids, with the latter consisting of two clearly defined subdomains, a protein kinase-like and a cyclase catalytic. Four potential N-linked glycosylation sites are present in the extracellular domain and 4 presumed phosphorylatable serine residues are conserved in the cyclase catalytic domain. Northern blot analysis demonstrated that the 4.5 kb mRNA for DsPTGC04 is expressed only in the testis. Antibodies raised against two synthetic peptides, ^<800>WVENPDERPN^<809> and ^<1080>KPPPQKLSAEVMEAAANREIPEDL^<1103>, corresponding to two carboxyl-terminal portions of DsPTGC04, reacted with a protein of about 120 kDa in D. setosum spermatozoa and testis, but not with any protein in the ovary, eggs, or intestine. Immunohistochemistry showed that both antibodies react with the mature spermatozoa in the testis.

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  • Zoological Science

    Zoological Science 18 (5), 687-693, 2001-07

    Zoological Society of Japan

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