1lfk
From Proteopedia
(New page: 200px<br /><applet load="1lfk" size="450" color="white" frame="true" align="right" spinBox="true" caption="1lfk, resolution 1.70Å" /> '''Crystal structure of...) |
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| - | [[Image:1lfk.gif|left|200px]]<br /><applet load="1lfk" size=" | + | [[Image:1lfk.gif|left|200px]]<br /><applet load="1lfk" size="350" color="white" frame="true" align="right" spinBox="true" |
caption="1lfk, resolution 1.70Å" /> | caption="1lfk, resolution 1.70Å" /> | ||
'''Crystal structure of OxyB, a Cytochrome P450 Implicated in an Oxidative Phenol Coupling Reaction During Vancomycin Biosynthesis'''<br /> | '''Crystal structure of OxyB, a Cytochrome P450 Implicated in an Oxidative Phenol Coupling Reaction During Vancomycin Biosynthesis'''<br /> | ||
==Overview== | ==Overview== | ||
| - | Gene-inactivation studies point to the involvement of OxyB in catalyzing | + | Gene-inactivation studies point to the involvement of OxyB in catalyzing the first oxidative phenol coupling reaction during glycopeptide antibiotic biosynthesis. The oxyB gene has been cloned and sequenced from the vancomycin producer Amycolatopsis orientalis, and the hemoprotein has been produced in Escherichia coli, crystallized, and its structure determined to 1.7-A resolution. OxyB gave UV-visible spectra characteristic of a P450-like hemoprotein in the low spin ferric state. After reduction to the ferrous state by dithionite or by spinach ferredoxin and ferredoxin reductase, the CO-ligated form gave a 450-nm peak in a UV-difference spectrum. Addition of putative heptapeptide substrates to resting OxyB produced type I changes to the UV spectrum, but no turnover was observed in the presence of ferredoxin and ferredoxin reductase, showing that either the peptides or the reduction system, or both, are insufficient to support a full catalytic cycle. OxyB exhibits the typical P450-fold, with helix L containing the signature sequence FGHGXHXCLG and Cys(347) being the proximal axial thiolate ligand of the heme iron. The structural similarity of OxyB is highest to P450nor, P450terp, CYP119, and P450eryF. In OxyB, the F and G helices are rotated out of the active site compared with P450nor, resulting in a much more open active site, consistent with the larger size of the presumed heptapeptide substrate. |
==About this Structure== | ==About this Structure== | ||
| - | 1LFK is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Amycolatopsis_orientalis Amycolatopsis orientalis] with HEM as [http://en.wikipedia.org/wiki/ligand ligand]. Full crystallographic information is available from [http:// | + | 1LFK is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Amycolatopsis_orientalis Amycolatopsis orientalis] with <scene name='pdbligand=HEM:'>HEM</scene> as [http://en.wikipedia.org/wiki/ligand ligand]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1LFK OCA]. |
==Reference== | ==Reference== | ||
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[[Category: Single protein]] | [[Category: Single protein]] | ||
[[Category: Pylypenko, O.]] | [[Category: Pylypenko, O.]] | ||
| - | [[Category: Robinson, J | + | [[Category: Robinson, J A.]] |
[[Category: Schlichting, I.]] | [[Category: Schlichting, I.]] | ||
[[Category: Vitali, F.]] | [[Category: Vitali, F.]] | ||
| - | [[Category: Vrijbloed, J | + | [[Category: Vrijbloed, J W.]] |
[[Category: Zerbe, K.]] | [[Category: Zerbe, K.]] | ||
[[Category: Zhang, W.]] | [[Category: Zhang, W.]] | ||
| Line 23: | Line 23: | ||
[[Category: oxidative phenol coupling reaction p450 vancomycin]] | [[Category: oxidative phenol coupling reaction p450 vancomycin]] | ||
| - | ''Page seeded by [http:// | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 13:44:36 2008'' |
Revision as of 11:44, 21 February 2008
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Crystal structure of OxyB, a Cytochrome P450 Implicated in an Oxidative Phenol Coupling Reaction During Vancomycin Biosynthesis
Overview
Gene-inactivation studies point to the involvement of OxyB in catalyzing the first oxidative phenol coupling reaction during glycopeptide antibiotic biosynthesis. The oxyB gene has been cloned and sequenced from the vancomycin producer Amycolatopsis orientalis, and the hemoprotein has been produced in Escherichia coli, crystallized, and its structure determined to 1.7-A resolution. OxyB gave UV-visible spectra characteristic of a P450-like hemoprotein in the low spin ferric state. After reduction to the ferrous state by dithionite or by spinach ferredoxin and ferredoxin reductase, the CO-ligated form gave a 450-nm peak in a UV-difference spectrum. Addition of putative heptapeptide substrates to resting OxyB produced type I changes to the UV spectrum, but no turnover was observed in the presence of ferredoxin and ferredoxin reductase, showing that either the peptides or the reduction system, or both, are insufficient to support a full catalytic cycle. OxyB exhibits the typical P450-fold, with helix L containing the signature sequence FGHGXHXCLG and Cys(347) being the proximal axial thiolate ligand of the heme iron. The structural similarity of OxyB is highest to P450nor, P450terp, CYP119, and P450eryF. In OxyB, the F and G helices are rotated out of the active site compared with P450nor, resulting in a much more open active site, consistent with the larger size of the presumed heptapeptide substrate.
About this Structure
1LFK is a Single protein structure of sequence from Amycolatopsis orientalis with as ligand. Full crystallographic information is available from OCA.
Reference
Crystal structure of OxyB, a cytochrome P450 implicated in an oxidative phenol coupling reaction during vancomycin biosynthesis., Zerbe K, Pylypenko O, Vitali F, Zhang W, Rouset S, Heck M, Vrijbloed JW, Bischoff D, Bister B, Sussmuth RD, Pelzer S, Wohlleben W, Robinson JA, Schlichting I, J Biol Chem. 2002 Dec 6;277(49):47476-85. Epub 2002 Aug 30. PMID:12207020
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