Species specificity of thimet oligopeptidase (EC 3.4.24.15).
The recombinant rat testes metallo-endooligopeptidase (EC 3.4.24.15) and the rabbit brain endooligopeptidase A (formerly EC 3.4.22.19) were compared, side-by-side, in view of their striking similarities in both the physicochemical features and the specificities for oligopeptides. Concerning the tissue distribution in rat and rabbit, no relation between the levels of enzyme activity in cytosol and the levels of metallo-endooligopeptidase 24.15 mRNA could be established. The results suggest that the predominant neuropeptide-metabolizing activity attributed to the metallo-endooligopeptidase 24.15 is performed by, at least, two distinct cytosolic enzymes, one predominant in rat testes and the other in rabbit brain and testes, and possibly also in rat brain. Both enzymes are activated by dithiothreitol and irreversibly inhibited by a SH-affinity labeling dynorphin-related compound, but they are not inhibited by EDTA in a concentration dependent manner. Both enzymes exhibit the same specificity toward several bioactive peptides, except for LH-RH and substance P, which are only hydrolysed by the rat testes enzyme. Taken together, these results lead us to conclude that it is unlikely that the recombinant rat testes metallo-endooligopeptidase 24.15 and the rabbit brain endooligopeptidase A are the same molecule although they might belong to the same family of oligopeptidases.
Hayashi
M A
MA
Laboratory of Biochemistry and Biophysics, Institute Butantan, São Paulo, Brazil.
Gomes
M D
MD
Rebouças
N A
NA
Fernandes
B L
BL
Ferro
E S
ES
de Camargo
A C
AC
D000595:Amino Acid Sequence; D000818:Animals; D015152:Blotting, Northern; D015153:Blotting, Western; D001921:Brain / Q000201:enzymology; D002614:Chelating Agents / Q000494:pharmacology; D003600:Cytosol / Q000201:enzymology; D004229:Dithiothreitol / Q000494:pharmacology; D004492:Edetic Acid / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D006868:Hydrolysis; D008297:Male; D008666:Metalloendopeptidases / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D008969:Molecular Sequence Data; D011817:Rabbits; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry; D013045:Species Specificity; D013379:Substrate Specificity; D013439:Sulfhydryl Reagents / Q000494:pharmacology; D013737:Testis / Q000201:enzymology
D002614:Chelating Agents; D011994:Recombinant Proteins; D013439:Sulfhydryl Reagents; D004492:Edetic Acid; D008666:Metalloendopeptidases; C022510:thimet oligopeptidase; D004229:Dithiothreitol