Crystal structure of carbapenem synthase (CarC)

被引:81
作者
Clifton, IJ
Doan, LX
Sleeman, MC
Topf, M
Suzuki, H
Wilmouth, RC
Schofield, CJ
机构
[1] Oxford Ctr Mol Sci, Oxford OX1 3QZ, England
[2] Dyson Perrins Lab, Oxford OX1 3QZ, England
[3] Phys & Theoret Chem Lab, Oxford OX1 3QZ, England
[4] Nanyang Technol Univ, Sch Biol Sci, Singapore 637616, Singapore
关键词
D O I
10.1074/jbc.M213054200
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 proposed biosynthetic pathway to the carbapenem antibiotics proceeds via epimerization/desaturation of a carbapenam in an unusual process catalyzed by an iron- and 2-oxoglutarate-dependent oxygenase, CarC. Crystal structures of CarC complexed with Fe(II) and 2-oxoglutarate reveal it to be hexameric (space group C2221), consistent with solution studies. CarC monomers contain a double-stranded beta-helix core that supports ligands binding a single Fe(II) to which 2-oxoglutarate complexes in a bi-dentate manner. A structure was obtained with L-N-acetylproline acting as a substrate analogue. Quantum mechanical/molecular mechanical modeling studies with stereoisomers of carbapenams and carbapenems were used to investigate substrate binding. The combined work will stimulate further mechanistic studies and aid in the engineering of carbapenem biosynthesis.
引用
收藏
页码:20843 / 20850
页数:8
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