Structure of a dinuclear iron cluster-containing β-hydroxylase active in antibiotic biosynthesis

Thomas M. Makris, Cory J. Knoot, Carrie M Wilmot, John D Lipscomb

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

A family of dinuclear iron cluster-containing oxygenases that catalyze β-hydroxylation tailoring reactions in natural product biosynthesis by nonribosomal peptide synthetase (NRPS) systems was recently described [Makris, T. M., Chakrabarti, M., Münck, E., and Lipscomb, J. D. (2010) Proc. Natl. Acad. Sci. U.S.A. 107, 15391-15396]. Here, the 2.17 Å X-ray crystal structure of the archetypal enzyme from the family, CmlA, is reported. CmlA catalyzes β-hydroxylation of l-p-aminophenylalanine during chloramphenicol biosynthesis. The fold of the N-terminal domain of CmlA is unlike any previously reported, but the C-terminal domain has the αββα fold of the metallo-β-lactamase (MBL) superfamily. The diiron cluster bound in the C-terminal domain is coordinated by an acetate, three His residues, two Asp residues, one Glu residue, and a bridging oxo moiety. One of the Asp ligands forms an unusual monodentate bridge. No other oxygen-activating diiron enzyme utilizes this ligation or the MBL protein fold. The N-terminal domain facilitates dimerization, but using computational docking and a sequence-based structural comparison to homologues, we hypothesize that it likely serves additional roles in NRPS recognition and the regulation of O2 activation.

Original languageEnglish (US)
Pages (from-to)6662-6671
Number of pages10
JournalBiochemistry
Volume52
Issue number38
DOIs
StatePublished - Sep 24 2013

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Peptide Synthases
Hydroxylation
Biosynthesis
Viperidae
Mixed Function Oxygenases
Iron
Anti-Bacterial Agents
Oxygenases
Dimerization
Chloramphenicol
Enzymes
Biological Products
Ligation
Acetates
Crystal structure
Chemical activation
X-Rays
Oxygen
Ligands
X rays

Cite this

Structure of a dinuclear iron cluster-containing β-hydroxylase active in antibiotic biosynthesis. / Makris, Thomas M.; Knoot, Cory J.; Wilmot, Carrie M; Lipscomb, John D.

In: Biochemistry, Vol. 52, No. 38, 24.09.2013, p. 6662-6671.

Research output: Contribution to journalArticle

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