Crystal Structure of Carbapenem Synthase (CarC)

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 hexame...

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Bibliographic Details
Published in:The Journal of biological chemistry 2003-06, Vol.278 (23), p.20843-20850
Main Authors: Clifton, Ian J., Doan, Linh X., Sleeman, Mark C., Topf, Maya, Suzuki, Hikokazu, Wilmouth, Rupert C., Schofield, Christopher J.
Format: Article
Language:English
Subjects:
Binding Sites
Carbapenems - biosynthesis
Crystallography
Iron - chemistry
Ketoglutaric Acids - chemistry
Oxidoreductases - chemistry
Oxidoreductases - metabolism
Pectobacterium carotovorum - enzymology
Protein Structure, Secondary
Protein Structure, Tertiary
Substrate Specificity
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Summary: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 β-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.
ISSN:0021-9258
1083-351X
DOI:10.1074/jbc.M213054200