4OJ8

Crystal structure of carbapenem synthase in complex with (3S,5S)-carbapenam

Summary for 4OJ8

• Entry DOI: 10.2210/pdb4oj8/pdb
• Descriptor: (5R)-carbapenem-3-carboxylate synthase, FE (II) ION, 2-OXOGLUTARIC ACID, ... (6 entities in total)
• Functional Keywords: oxidoreductase, oxygenase, iron, 2-oxoglutaric acid, antibiotic biosynthesis, carbapenam
• Biological source: Pectobacterium carotovorum subsp. carotovorum
• Cellular location: Cytoplasm : Q9XB59
• Total number of polymer chains: 3
• Total formula weight: 102471.87

Authors

Boal, A.K.,Rosenzweig, A.C. (deposition date: 2014-01-20, release date: 2014-04-02, Last modification date: 2023-09-20)

Primary citation

Chang, W.C.,Guo, Y.,Wang, C.,Butch, S.E.,Rosenzweig, A.C.,Boal, A.K.,Krebs, C.,Bollinger, J.M.
Mechanism of the C5 stereoinversion reaction in the biosynthesis of carbapenem antibiotics.
Science, 343:1140-1144, 2014

PubMed Abstract: The bicyclic β-lactam/2-pyrrolidine precursor to all carbapenem antibiotics is biosynthesized by attachment of a carboxymethylene unit to C5 of L-proline followed by β-lactam ring closure. Carbapenem synthase (CarC), an Fe(II) and 2-(oxo)glutarate (Fe/2OG)-dependent oxygenase, then inverts the C5 configuration. Here we report the structure of CarC in complex with its substrate and biophysical dissection of its reaction to reveal the stereoinversion mechanism. An Fe(IV)-oxo intermediate abstracts the hydrogen (H•) from C5, and tyrosine 165, a residue not visualized in the published structures of CarC lacking bound substrate, donates H• to the opposite face of the resultant radical. The reaction oxidizes the Fe(II) cofactor to Fe(III), limiting wild-type CarC to one turnover, but substitution of the H•-donating tyrosine disables stereoinversion and confers to CarC the capacity for catalytic substrate oxidation.

PubMed: 24604200
DOI: 10.1126/science.1248000

Experimental method

X-RAY DIFFRACTION (2.1 Å)