Catalogo Articoli (Spogli Riviste)

OPAC HELP

Titolo:
Identification and designing of the S3 site of aqualysin I, a thermophilicsubtilisin-related serine protease
Autore:
Tanaka, T; Matsuzawa, H; Ohta, T;
Indirizzi:
Univ Tokyo, Dept Biotechnol, Bunkyo Ku, Tokyo 1138657, Japan Univ Tokyo Tokyo Japan 1138657 otechnol, Bunkyo Ku, Tokyo 1138657, Japan
Titolo Testata:
JOURNAL OF BIOCHEMISTRY
fascicolo: 6, volume: 125, anno: 1999,
pagine: 1016 - 1021
SICI:
0021-924X(199906)125:6<1016:IADOTS>2.0.ZU;2-B
Fonte:
ISI
Lingua:
ENG
Soggetto:
THERMUS-AQUATICUS YT-1; PROTEINASE-K; SUBSTRATE-SPECIFICITY; ESCHERICHIA-COLI; EGLIN-C; 3-DIMENSIONAL STRUCTURE; DIRECTED MUTAGENESIS; INHIBITOR COMPLEXES; CRYSTAL-STRUCTURE; DISULFIDE BONDS;
Keywords:
aqualysin I; P3-specificity; S3 subsite; serine protease; subtilsin;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
46
Recensione:
Indirizzi per estratti:
Indirizzo: Tanaka, T Toyohashio,niv Technol, Dept Ecol Engn, Div Biosci & Biotechnol,Tempaku Ch Toyohashi Univ Technol Tempaku Cho Aichi Japan 4418580 mpaku Ch
Citazione:
T. Tanaka et al., "Identification and designing of the S3 site of aqualysin I, a thermophilicsubtilisin-related serine protease", J BIOCHEM, 125(6), 1999, pp. 1016-1021

Abstract

Aqualysin I is a bacterial subtilisin-related alkaline serine protease, originating in Thermus aquaticus YT-1, Based on computational analysis, we predicted that two residues, Ser(102) and Gly(131), form the 53 site of aqualysin I, and we proved that this prediction by site-directed mutagenesis. Toalter the P3-specificity of the enzyme, we built a "wall" on the S3 site edge by introducing a bulky side chain at target sites. Six mutant proteins were prepared: S102H, S102K, S102E, G131H, G131K, and G;131D, The mutant enzymes were examined with two kinetically typical peptides for aqualysin I, suc-X-Ala-Ala-pNA, where X is Ala or Phe, All mutations reduced the efficiency for the Phe-containing peptide, while they raised the k(cat), values for the Ala-containing peptide. Especially, the S102K mutant protein hydrolyzed the polyalanine peptide efficiently. The strategies we have adopted in this paper are applicable to all subtilisin-related enzymes.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 03/04/20 alle ore 04:25:04