engleski

Characterization of a membrane-bound angiotensin- converting enzyme isoform in crayfish testis and evidence for its release into the seminal fluid

In the present study, an isoform of angiotensin-converting enzyme was characterized from the testis of a decapod crustacean, the crayfish Astacus leptodactylus. Angiotensin- converting enzyme cDNA, obtained by 3′- to 5′ RACE of testis RNAs, codes for a predicted one-domain protein similar to the mammalian germinal isoform of angiotensin- converting enzyme. All amino acid residues involved in enzyme activity are highly conserved, and a potential C- terminus transmembrane anchor may be predicted from the sequence. Comparison of this testicular isoform with angiotensin-converting enzyme from other crustaceans, namely Carcinus maenas, Homarus americanus (both reconstituted for this study from expressed-sequence tag data) and Daphnia pulex, suggests that membrane-bound angiotensin-converting enzyme occurs widely in crustaceans, conversely to other invertebrate groups where angiotensin-converting enzyme is predominantly a soluble protein. In situ hybridization and immunohistochemistry performed on testis sections show that angiotensin- converting enzyme mRNA is mainly localized in spermatogonias, whereas protein is present in spermatozoids. By contrast, in vas deferens, immunoreactivity is detected in the seminal fluid rather than in germ cells. Accordingly, angiotensin-converting enzyme activity assays of testis and vas deferens extracts demonstrate that the enzyme is present in the membrane fraction in testis, but in the soluble fraction in vas deferens. Taken together, the results obtained in the present study suggest that, during the migration of spermatozoids from testis to vas deferens, the enzyme is cleaved from the membrane of the germ cells and released into the seminal fluid. To our knowledge, this present study is the first to report such a maturation process for angiotensin-converting enzyme outside of mammals.

angiotensin-converting enzyme ; crayfish ;

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