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1qjf

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[[Category: sulfoxide]]
[[Category: sulfoxide]]
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Revision as of 14:55, 5 November 2007


1qjf, resolution 1.40Å

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ISOPENICILLIN N SYNTHASE FROM ASPERGILLUS NIDULANS (MONOCYCLIC SULFOXIDE-FE COMPLEX)

Overview

Isopenicillin N synthase (IPNS), a non-haem iron-dependent oxidase, catalyses the biosynthesis of isopenicillin N (IPN), the precursor of all, penicillins and cephalosporins. The key steps in this reaction are the two, iron-dioxygen-mediated ring closures of the tripeptide, delta-(L-alpha-aminoadipoyl)-L-cysteinyl-D-valine (ACV). It has been, proposed that the four-membered beta-lactam ring forms initially, associated with a highly oxidized iron(iv)-oxo (ferryl) moiety, which, subsequently mediates closure of the five-membered thiazolidine ring. Here, we describe observation of the IPNS reaction in crystals by X-ray, crystallography. IPNS Fe2+ substrate crystals were grown anaerobically, exposed to high pressures of oxygen to promote reaction and frozen, and, their structures were elucidated by X-ray diffraction. Using the natural, substrate ACV, this resulted in the IPNS x Fe2+ x IPN product complex., With the substrate analogue, delta-(L-alpha-aminoadipoyl)-L-cysteinyl-L-S-methylcysteine (ACmC) in the, crystal, the reaction cycle was interrupted at the monocyclic stage. These, mono- and bicyclic structures support our hypothesis of a two-stage, reaction sequence leading to penicillin. Furthermore, the formation of a, monocyclic sulphoxide product from ACmC is most simply explained by the, interception of a high-valency iron-oxo species.

About this Structure

1QJF is a Single protein structure of sequence from Emericella nidulans with SO4, FE2 and ACS as ligands. Structure known Active Site: SA. Full crystallographic information is available from OCA.

Reference

The reaction cycle of isopenicillin N synthase observed by X-ray diffraction., Burzlaff NI, Rutledge PJ, Clifton IJ, Hensgens CM, Pickford M, Adlington RM, Roach PL, Baldwin JE, Nature. 1999 Oct 14;401(6754):721-4. PMID:10537113

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