Identification of reactive conserved histidines in phosphoenolpyruvate carboxykinases from Escherichia coli and Saccharomyces cerevisiae

被引:7
作者
Bazaes, S
Montecinos, L
Krautwurst, H
Goldie, H
Cardemil, E
Jabalquinto, AM
机构
[1] UNIV SANTIAGO CHILE,FAC QUIM & BIOL,DEPT CIENCIAS QUIM,SANTIAGO 2,CHILE
[2] UNIV METROPOLITANA CIENCIAS EDUC,DEPT QUIM,SANTIAGO,CHILE
[3] UNIV SASKATCHEWAN,DEPT MICROBIOL,SASKATOON,SK S7N 0W0,CANADA
来源
BIOCHIMICA ET BIOPHYSICA ACTA-PROTEIN STRUCTURE AND MOLECULAR ENZYMOLOGY | 1997年 / 1337卷 / 02期
关键词
phosphoenolpyruvate carboxykinase; histidyl residue; inactivation; (E-coli); (S-cerevisiae);
D O I
10.1016/S0167-4838(96)00155-0
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
Escherichia coli and Saccharomyces cerevisiae phosphoenolpyruvate (PEP) carboxykinases are inactivated by diethylpyrocarbonate (DEP). Inactivation follows pseudo-first-order kinetics and exhibits a second order rate constant of 0.8 M(-1) s(-1) for the bacterial enzyme and of 3.3 M(-1) s(-1) for the yeast carboxykinase, A mixture of ADP + PEP + MnCl2 protects against inactivation by DEP, suggesting that residues within the active site are being modified. After digestion of the modified proteins with trypsin, the labeled peptides were isolated by reverse-phase high-performance liquid chromatography and sequenced by Edman degradation. His-271 of E. coli carboxykinase and His-273 of the yeast enzyme were identified as the reactive amino-acid residues. The modified histidine residues occupy equivalent positions in these enzymes, and they are located in a highly conserved region of all ATP-dependent phosphoenolpyruvate carboxykinases described so far.
引用
收藏
页码:166 / 174
页数:9
相关论文
共 41 条
[31]  
ROJAS MC, 1993, BIOCHIM BIOPHYS ACTA, V1164, P143
[32]   STEREOCHEMICAL COURSE OF THIOPHOSPHORYL GROUP TRANSFER CATALYZED BY MITOCHONDRIAL PHOSPHOENOLPYRUVATE CARBOXYKINASE [J].
SHEU, KF ;
HO, HT ;
NOLAN, LD ;
MARKOVITZ, P ;
RICHARD, JP ;
UTTER, MF ;
FREY, PA .
BIOCHEMISTRY, 1984, 23 (08) :1779-1783
[33]   Snapshot of an enzyme reaction intermediate in the structure of the ATP-Mg2+-oxalate ternary complex of Escherichia coli PEP carboxykinase [J].
Tari, LW ;
Matte, A ;
Pugazhenthi, U ;
Goldie, H ;
Delbaere, LTJ .
NATURE STRUCTURAL BIOLOGY, 1996, 3 (04) :355-363
[34]   PURIFICATION OF PHOSPHOENOLPYRUVATE CARBOXYKINASE FROM SACCHAROMYCES-CEREVISIAE AND ITS USE FOR BICARBONATE ASSAY [J].
TORTORA, P ;
HANOZET, GM ;
GUERRITORE, A .
ANALYTICAL BIOCHEMISTRY, 1985, 144 (01) :179-185
[35]   THE FUNCTIONS AND CONSENSUS MOTIFS OF 9 TYPES OF PEPTIDE SEGMENTS THAT FORM DIFFERENT TYPES OF NUCLEOTIDE-BINDING SITES [J].
TRAUT, TW .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1994, 222 (01) :9-19
[36]  
TSOU C, 1962, SCI SINICA, V11, P1535
[38]   INHIBITION OF PHOSPHOENOLPYRUVATE CARBOXYKINASE FROM TRYPANOSOMA-(SCHIZOTRYPANUM)-CRUZI EPIMASTIGOTES BY 3-MERCAPTOPICOLINIC ACID - INVITRO AND INVIVO STUDIES [J].
URBINA, JA ;
OSORNO, CE ;
ROJAS, A .
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1990, 282 (01) :91-99
[39]  
Utter M.F., 1972, Enzymes, V6, P117, DOI [10.1016/S1874-6047(08)60039-6, DOI 10.1016/S1874-6047(08)60039-6]
[40]   SINGLE ACTIVE-SITE HISTIDINE IN D-XYLOSE ISOMERASE FROM STREPTOMYCES-VIOLACEORUBER - IDENTIFICATION BY CHEMICAL DERIVATIZATION AND PEPTIDE-MAPPING [J].
VANGRYSPERRE, W ;
AMPE, C ;
KERSTERSHILDERSON, H ;
TEMPST, P .
BIOCHEMICAL JOURNAL, 1989, 263 (01) :195-199