Site-directed mutagenesis of Flaveria trinervia phosphoenolpyruvate carboxylase: Arg(450) and Arg(767) are essential for catalytic activity and Lys(829) affects substrate binding

被引：12
作者：
Gao, Y [1 ]
Woo, KC [1 ]
机构：
[1] NO TERR UNIV,SCH BIOL SCI,DARWIN,NT 0909,AUSTRALIA
来源：
FEBS LETTERS | 1996年 / 392卷 / 03期
关键词：
phosphoenolpyruvate carboxylase; arginine; lysine; substrate binding; Flaveria trinervia;
D O I：
10.1016/0014-5793(96)00832-0
中图分类号：
Q5 [生物化学]; Q7 [分子生物学];
学科分类号：
071010 ; 081704 ;
摘要：
Phosphoenolpyruvate carboxylases (PEPC) of all known sequences contain 11 conserved arginine and two lysine residues located in highly conservative regions. Previous chemical modifications show that arginine and lysine residues are essential for catalytic activity. Three conserved residues, Arg(450), Arg(767) and Lys(829) in PEPC of Flaveria trinervia were converted to glycine. All three mutant PEPC proteins were similarly expressed in Escherichia coli, However, mutant Gly(450) and Gly(767) PEPCs had no catalytic activity and Gly(829) PEPC showed increased K-m for PEP and Mg2+. It seems that Arg(450) and Arg(767) are essential for PEPC function while Lys(829) might be associated with PEP and/or Mg2+ binding domain.
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收藏
页码：285 / 288
页数：4
相关论文
共 25 条
[1]   CHEMICAL MODIFICATION OF THE PHOSPHOENOLPYRUVATE CARBOXYLASE FROM MAIZE LEAVES AND ITS CONFORMATION IN ISOTROPIC SOLUTION - STUDIES VIA TRIPLET LIFETIME AND ROTATIONAL DIFFUSION USING EOSIN ISOTHIOCYANATE AS LABEL [J].
ANDREO, CS ;
IGLESIAS, AA ;
PODESTA, FE ;
WAGNER, R .
BIOCHIMICA ET BIOPHYSICA ACTA, 1986, 870 (02) :292-301
[2]   PROPERTIES OF AN ESCHERICHIA-COLI MUTANT DEFICIENT IN PHOSPHOENOLPYRUVATE CARBOXYLASE CATALYTIC ACTIVITY [J].
COOMES, MW ;
MITCHELL, BK ;
BEEZLEY, A ;
SMITH, TE .
JOURNAL OF BACTERIOLOGY, 1985, 164 (02) :646-652
[3]   NODULE PHOSPHOENOLPYRUVATE CARBOXYLASE - A REVIEW [J].
DEROCHE, ME ;
CARRAYOL, E .
PHYSIOLOGIA PLANTARUM, 1988, 74 (04) :775-782
[4]   SITE-DIRECTED MUTAGENESIS OF LYS(600) IN PHOSPHOENOLPYRUVATE CARBOXYLASE OF FLAVERIA-TRINERVIA - ITS ROLES IN CATALYTIC AND REGULATORY FUNCTIONS [J].
GAO, Y ;
WOO, KC .
FEBS LETTERS, 1995, 375 (1-2) :95-98
[5]   Regulation of phosphoenolpyruvate carboxylase in Zea mays by protein phosphorylation and metabolites and their roles in photosynthesis [J].
Gao, Y ;
Woo, KC .
AUSTRALIAN JOURNAL OF PLANT PHYSIOLOGY, 1996, 23 (01) :25-32
[6]   C-4 PHOTOSYNTHESIS - A UNIQUE BLEND OF MODIFIED BIOCHEMISTRY, ANATOMY AND ULTRASTRUCTURE [J].
HATCH, MD .
BIOCHIMICA ET BIOPHYSICA ACTA, 1987, 895 (02) :81-106
[7]   THE PRESENCE OF ESSENTIAL HISTIDINE-RESIDUES IN PHOSPHOENOLPYRUVATE CARBOXYLASE FROM MAIZE LEAVES [J].
IGLESIAS, AA ;
ANDREO, CS .
BIOCHIMICA ET BIOPHYSICA ACTA, 1983, 749 (01) :9-17
[8]   INACTIVATION OF PHOSPHOENOLPYRUVATE CARBOXYLASE FROM MAIZE LEAVES BY MODIFICATION WITH PHENYLGLYOXAL [J].
IGLESIAS, AA ;
GONZALEZ, DH ;
ANDREO, CS .
BIOCHIMICA ET BIOPHYSICA ACTA, 1984, 788 (01) :41-47
[9]   ISOLATION AND SEQUENCE OF AN ACTIVE-SITE PEPTIDE FROM MAIZE LEAF PHOSPHOENOLPYRUVATE CARBOXYLASE INACTIVATED BY PYRIDOXAL 5'-PHOSPHATE [J].
JIAO, JA ;
PODESTA, FE ;
CHOLLET, R ;
OLEARY, MH ;
ANDREO, CS .
BIOCHIMICA ET BIOPHYSICA ACTA, 1990, 1041 (03) :291-295
[10]   SORGHUM PHOSPHOENOLPYRUVATE CARBOXYLASE GENE FAMILY - STRUCTURE, FUNCTION AND MOLECULAR EVOLUTION [J].
LEPINIEC, L ;
KERYER, E ;
PHILIPPE, H ;
GADAL, P ;
CRETIN, C .
PLANT MOLECULAR BIOLOGY, 1993, 21 (03) :487-502
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