1qje
From Proteopedia
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==Overview== | ==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 ... | + | 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== | ==About this Structure== | ||
| - | 1QJE is a | + | 1QJE is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Emericella_nidulans Emericella nidulans] with SO4, FE2 and IP1 as [http://en.wikipedia.org/wiki/ligands ligands]. Structure known Active Site: SA. Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=1QJE OCA]. |
==Reference== | ==Reference== | ||
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[[Category: penicillin biosynthesis]] | [[Category: penicillin biosynthesis]] | ||
| - | ''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on | + | ''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on Mon Nov 5 14:03:29 2007'' |
Revision as of 11:58, 5 November 2007
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ISOPENICILLIN N SYNTHASE FROM ASPERGILLUS NIDULANS (IP1-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
1QJE is a Single protein structure of sequence from Emericella nidulans with SO4, FE2 and IP1 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
Page seeded by OCA on Mon Nov 5 14:03:29 2007
