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Titolo:
PUTATIVE FUNCTIONS OF PHENYLALANINE-350 OF PSEUDOMONAS-PUTIDA CYTOCHROME P-450(CAM)
Autore:
YASUKOCHI T; OKADA O; HARA T; SAGARA Y; SEKIMIZU K; HORIUCHI T;
Indirizzi:
KYUSHU UNIV,FAC PHARMACEUT SCI,DEPT MICROBIOL,HIGASHI KU FUKUOKA 812 JAPAN KYUSHU UNIV,FAC PHARMACEUT SCI,DEPT MICROBIOL,HIGASHI KU FUKUOKA 812 JAPAN NAKAMURA GAKUEN COLL,DEPT FOOD & NUTR,JONAN KU FUKUOKA 81401 JAPAN SOKA UNIV,FAC ENGN,DEPT BIOENGN HACHIOJI TOKYO 192 JAPAN
Titolo Testata:
Biochimica et biophysica acta. Protein structure and molecular enzymology
fascicolo: 1, volume: 1204, anno: 1994,
pagine: 84 - 90
SICI:
0167-4838(1994)1204:1<84:PFOPOP>2.0.ZU;2-S
Fonte:
ISI
Lingua:
ENG
Soggetto:
ELECTRON-TRANSFER COMPLEX; CRYSTAL-STRUCTURE; PUTIDAREDOXIN REDUCTASE; NUCLEOTIDE-SEQUENCES; AMINO-ACID; MUTAGENESIS; P-450CAM; PROTEINS; CLONING; HYDROXYLASE;
Keywords:
CYTOCHROME P-450(CAM); MUTANT; SITE-DIRECTED MUTAGENESIS; ELECTRON TRANSFER; (PSEUDOMONAS PUTIDA);
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Science Citation Index Expanded
Citazioni:
39
Recensione:
Indirizzi per estratti:
Citazione:
T. Yasukochi et al., "PUTATIVE FUNCTIONS OF PHENYLALANINE-350 OF PSEUDOMONAS-PUTIDA CYTOCHROME P-450(CAM)", Biochimica et biophysica acta. Protein structure and molecular enzymology, 1204(1), 1994, pp. 84-90

Abstract

Cytochrome P-450(cam) hydroxylates d-camphor, using molecular oxygen and reducing equivalents transferred via putidaredoxin. We constructedmutant genes in which Phe-350 of P-450(cam) was replaced by Leu, Tyr,or His by site-directed mutagenesis, expressed them in Escherichia coli, purified the mutant proteins, and compared their enzymic properties with those of the wild type P-450(cam). NADH oxidation rate of the Tyr mutant in the reconstituted system with putidaredoxin and putidaredoxin reductase was similar to that of the wild type enzyme, while the Leu mutant and the His mutant showed 67% and 17% activity of that of the wild type, respectively. The affinities of these mutant proteins for camphor and the oxidized form of putidaredoxin were much the same asthose of the wild type protein. Rate constants for the reduction reaction of P-450(cam) by reduced putidaredoxin, a physiological electron donor for P-450(cam), of Tyr and His mutants were much the same as that of the wild type enzyme, whereas the Leu mutant showed approx. half that of the wild type. Thus, the aromatic ring of Phe-350 of P-450(cam) probably contributes to enhancing efficiency of the electron transfer yet does not seem to be essential for the reaction.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 05/07/20 alle ore 13:31:56