Catalogo Articoli (Spogli Riviste)

OPAC HELP

Titolo:
5-ENOLPYRUVYLSHIKIMATE-3-PHOSPHATE SYNTHASE OF BACILLUS-SUBTILIS IS AN ALLOSTERIC ENZYME - ANALYSIS OF ARG24-]ASP, PRO105-]SER AND HIS385-]LYS MUTATIONS SUGGESTS A HIDDEN PHOSPHOENOLPYRUVATE-BINDING SITE
Autore:
MAJUMDER K; SELVAPANDIYAN A; FATTAH FA; ARORA N; AHMAD S; BHATNAGAR RK;
Indirizzi:
INT CTR GENET ENGN & BIOTECHNOL,NII CAMPUS ARUNA ASAF ALI MARG NEW DELHI 110067 INDIA INT CTR GENET ENGN & BIOTECHNOL NEW DELHI 110067 INDIA
Titolo Testata:
European journal of biochemistry
fascicolo: 1, volume: 229, anno: 1995,
pagine: 99 - 106
SICI:
0014-2956(1995)229:1<99:5SOBIA>2.0.ZU;2-P
Fonte:
ISI
Lingua:
ENG
Soggetto:
AMINO-ACID BIOSYNTHESIS; ESCHERICHIA-COLI; AROA GENE; KLEBSIELLA-PNEUMONIAE; 3-PHOSPHATE SYNTHASE; HERBICIDE GLYPHOSATE; DIRECTED MUTAGENESIS; NUCLEOTIDE-SEQUENCE; ACTIVE-SITE; TRANSCARBAMOYLASE;
Keywords:
5-ENOLPYRUVYLSHIKIMATE-3-PHOSPHATE SYNTHASE; SITE-DIRECTED MUTAGENESIS; ALLOSTERIC ENZYME; GLYPHOSATE;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
32
Recensione:
Indirizzi per estratti:
Citazione:
K. Majumder et al., "5-ENOLPYRUVYLSHIKIMATE-3-PHOSPHATE SYNTHASE OF BACILLUS-SUBTILIS IS AN ALLOSTERIC ENZYME - ANALYSIS OF ARG24-]ASP, PRO105-]SER AND HIS385-]LYS MUTATIONS SUGGESTS A HIDDEN PHOSPHOENOLPYRUVATE-BINDING SITE", European journal of biochemistry, 229(1), 1995, pp. 99-106

Abstract

5-Enolpyruvylshikimate-3-phosphate synthase of Bacillus subtilis has been cloned, expressed and purified to near homogeneity. Clustal alignment of the amino acid sequences from different bacteria revealed several conserved residues located in the N-terminal, middle and C-terminal domains. The role of conserved Arg24, Pro105, and His385 residues has been examined by site-directed mutagenesis. Steady-state kinetic analysis of the native synthase exhibited allosteric behaviour, a featurethought to be unique amongst bacterial and plant 5-enolpyruvylshikimate-3-phosphate synthase enzymes investigated so far. Both substrates, phosphoenolpyruvate (P-pyruvate) and shikimate 3-phosphate have multiple interaction sites. There are two sites for P-pyruvate binding, catalytic and non-catalytic. Glyphosate (N-phosphonomethyl glycine) competes for binding at the catalytic site and does not interact at the secondary site. Glyphosate in the absence of ammonium ions increases cooperativity of P-pyruvate binding and favors dimerization of the enzyme through an interaction between P-pyruvate-binding sites. The ammonium-ion-activated 5-enolpyruvylshikimate-3-phosphate synthase displays no cooperativity with respect to P-pyruvate. Absence of ammonium ions decreases affinity for substrates and introduces cooperativity. Cooperativity was also introduced in the enzyme by point mutations, Arg24-->Asp and His385-->Lys. The latter mutant of the native enzyme exists as a dimer and aggregates to a tetrameric form in the presence of glyphosate. The occurrence of multimeric forms of the synthase has been demonstrated by staining for the enzyme activity on the native gel and by resolving purified enzyme preparations on a sucrose density gradient. A model describing the alteration in the aggregation status of the enzyme by the inhibitor, activator and the substrates has been proposed.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 19/09/20 alle ore 10:50:49