CMP kinase from Escherichia coli is structurally related to other nucleoside monophosphate kinases

被引:64
作者
Bucurenci, N
Sakamoto, H
Briozzo, P
Palibroda, N
Serina, L
Sarfati, RS
Labesse, G
Briand, G
Danchin, A
Barzu, O
Gilles, AM
机构
[1] INST PASTEUR,UNITE BIOCHIM REGULAT CELLULAIRES,F-75724 PARIS 15,FRANCE
[2] INST PASTEUR,UNITE CHIM ORGAN,F-75724 PARIS 15,FRANCE
[3] INST PASTEUR,UNITE REGULAT EXPRESS GENET,F-75724 PARIS 15,FRANCE
[4] UNIV PARIS 06,LAB MINERAL CRISTALLOG,F-75252 PARIS 05,FRANCE
[5] UNIV PARIS 07,F-75252 PARIS 05,FRANCE
[6] CNRS,UMR 9920,LAB BIOL STRUCT,F-91198 GIF SUR YVETTE,FRANCE
[7] UNIV LILLE 2,LAB APPLICAT SPECTROMETRIE MASSE,F-59405 LILLE,FRANCE
关键词
D O I
10.1074/jbc.271.5.2856
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
CMP kinase from Escherichia coli is a monomeric protein of 225 amino acid residues. The protein exhibits little overall sequence similarities with other known NMP kinases. However, residues involved in binding of substrates and/or in catalysis were found conserved, and sequence comparison suggested conservation of the global fold found in adenylate kinases or in several CMP/UMP kinases. The enzyme was purified to homogeneity, crystallized, and analyzed for its structural and catalytic properties. The crystals belong to the hexagonal space group P6(3), have unit cell parameters a = b = 82.3 Angstrom and c = 60.7 Angstrom and diffract x-rays to a 1.9 Angstrom resolution. The bacterial enzyme exhibits a fluorescence emission spectrum with maximum at 328 nm upon excitation at 295 nm, which suggests that the single tryptophan residue (Trp(30)) is located in a hydrophobic environment. Substrate specificity studies showed that CMP kinase from E. coli is active with ATP, dATP, or GTP as donors and with CMP, dCMP, and arabinofuranosyl-CMP as accepters. This is in contrast with CMP/UMP kinase from Dictyostelium discoideum, an enzyme active on CMP or UMP but much less active on the corresponding deoxynucleotides. Binding of CMP enhanced the affinity of E. coli CMP kinase for ATP or ADP, a particularity never described in this family of proteins that might explain inhibition of enzyme activity by excess of nucleoside monophosphate.
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页码:2856 / 2862
页数:7
相关论文
共 44 条
[1]  
ALTSCHUL SF, 1990, J MOL BIOL, V215, P403, DOI 10.1006/jmbi.1990.9999
[2]  
Anderson E. P., 1973, ENZYMES, V8, P49
[3]  
[Anonymous], ACTA CRYSTALLOGR D
[4]  
[Anonymous], 1973, GROUP TRANSFER A
[5]  
[Anonymous], LIFE SCI ADV
[6]   SIMPLE AND FAST PURIFICATION OF ESCHERICHIA-COLI ADENYLATE KINASE [J].
BARZU, O ;
MICHELSON, S .
FEBS LETTERS, 1983, 153 (02) :280-284
[7]  
BARZU O, 1993, ANN I PASTEUR ACTUAL, V4, P121
[8]   SALMONELLA-TYPHIMURIUM MUTANTS DEFECTIVE IN CYTIDINE MONOPHOSPHATE KINASE (CMK) [J].
BECK, CF ;
NEUHARD, J ;
THOMASSEN, E ;
INGRAHAM, JL ;
KLEKER, E .
JOURNAL OF BACTERIOLOGY, 1974, 120 (03) :1370-1379
[9]  
BELL JE, 1988, PROTEINS ENZYMES, P370
[10]  
BLINKLEY JP, 1986, J BACTERIOL, V168, P1457