Artigo
Basic aminopeptidase from rabbit kidney: Purification and partial characterization
Fecha
1996-11-01Registro en:
Brazilian Journal Of Medical And Biological Research. Sao Paulo: Assoc Bras Divulg Cientifica, v. 29, n. 11, p. 1437-1439, 1996.
0100-879X
WOS:A1996VR61500004
Autor
Oliveira, S. M.
Freitas, J. O.
Alves, Kaethy Bisan [UNIFESP]
Institución
Resumen
The aminopeptidase activity of a homogenate of rabbit kidney treated with Triton X-100 was measured using L-aminoacyl-2-naphthylamides (AA-NA). After gradient elution ion-exchange chromatography, four peaks of aminopeptidase activity were eluted. The enzyme eluted at 450 mu S containing 33.5% of the activity towards Arg-NA was applied to a Superdex 75 column and presented only one protein band on 10% SDS-polyacrylamide gel electrophoresis. This enzyme has an apparent molecular mass of 78 kDa, is five-fold activated by 0.15 M NaCl and the highest V-max/K-M ratio was obtained with Arg-NA. Enzyme activity was inhibited 100% by 0.13 mM sodium p-hydroxymercuribenzoate, 20% by 0.75 mM EDTA and 100% by 0.66 mM o-phenanthroline. Puromycin and bestatin behaved like competitive inhibitors with a K-i of 0.60 mM and 5.0 mu M, respectively.