Structure and function of the dihydropteroate synthase from Staphylococcus aureus

被引:150
作者
Hampele, IC [1 ]
DArcy, A [1 ]
Dale, GE [1 ]
Kostrewa, D [1 ]
Nielsen, J [1 ]
Oefner, C [1 ]
Page, MGP [1 ]
Schonfeld, HJ [1 ]
Stuber, D [1 ]
Then, RL [1 ]
机构
[1] F HOFFMANN LA ROCHE & CO LTD, DEPT PHARMA PRECLIN RES, CH-4070 BASEL, SWITZERLAND
关键词
DHPS; kinetics; sulfonamide-resistance; X-ray structure; S-aureus;
D O I
10.1006/jmbi.1997.0944
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
The gene encoding the dihydropteroate synthase of staphylococcus aureus has been cloned, sequenced and expressed in Escherichia coli. The protein has been purified for biochemical characterization and X-ray crystallographic studies. The enzyme is a dimer in solution, has a steady state kinetic mechanism that suggests random binding of the two substrates and half-site reactivity. The crystal structure of ape-enzyme and a binary complex with the substrate analogue hydroxymethylpterin pyrophosphate were determined at 2.2 Angstrom and 2.4 Angstrom resolution, respectively. The enzyme belongs to the group of ''TIM-barrel'' proteins and crystallizes as a non-crystallographic dimer. Only one molecule of the substrate analogue bound per dimer in the crystal. Sequencing of nine sulfonamide-resistant clinical isolates has shown that as many as 14 residues could be involved in resistance development. The residues are distributed over the surface of the protein, which defies a simple interpretation of their roles in resistance. Nevertheless, the three-dimensional structure of the substrate analogue binary complex could give important insight into the molecular mechanism of this enzyme. (C) 1997 Academic Press Limited.
引用
收藏
页码:21 / 30
页数:10
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