[Purification and physico-chemical properties of soluble carboxypeptidase N from cat brain gray matter].
Using affinity chromatography on diasorb-L-arginine and polyacrylamide gel electrophoresis, soluble carboxypeptidase H (E. C. 3.4.17.10) has been isolated from cat brain cortex and purified 598-fold with a 16% yield. The enzyme has a molecular mass of 50 kDa, consists of one polypeptide chain, and displays the maximum activity at pH 5.6. Carboxypeptidase H is a thiol-dependent metalloenzyme and contains a Zn2+ ion in its active center. The Km and V values for dansyl-Phe-Leu-Arg are 100 +/- 5 microM and 12.5 +/- 1.4 microM/min/mg of protein, respectively. The existence of two forms of soluble carboxypeptidase differing in isoelectric points and pH optima has been demonstrated. The enzyme with a pI of 4.8 has a pH optimum at 5.5-5.6, while that with a pI of 5.25-at 6.0.
Vernigora
A N
AN
Gengin
M T
MT
Nikishin
N N
NN
D000818:Animals; D001665:Binding Sites; D001921:Brain / Q000201:enzymology*; D002412:Cations; D002415:Cats; D002846:Chromatography, Affinity; D002855:Chromatography, Thin Layer; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D007526:Isoelectric Point; D001121:Lysine Carboxypeptidase / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D008670:Metals; D012995:Solubility
D002412:Cations; D008670:Metals; D001121:Lysine Carboxypeptidase
Ochistka i fiziko-khimicheskie svoĭstva rastvorimoĭ karboksipeptidazyN iz serogo veshchestva golovnogo mozga koshki.