PDBsum entry 1lgo

Biosynthetic protein

PDB id: 1lgo

Contents

Protein chain

155 a.a.

Theoretical model

PDB id:

1lgo

Name:

Biosynthetic protein

Title:

Structure of ethanolamine utilization protein eutk [precursor]

Structure:

Ethanolamine utilization protein. Chain: a. Synonym: putative carboxysome structural protein

Source:

Salmonella typhimurium. Bacteria

Authors:

R.Sagajkar,R.Ramchandra,A.Muthuvel

Key ref:

K.Zerbe et al. (2002). Crystal structure of OxyB, a cytochrome P450 implicated in an oxidative phenol coupling reaction during vancomycin biosynthesis. J Biol Chem, 277, 47476-47485. PubMed id: 12207020 DOI: 10.1074/jbc.M206342200

Date:

16-Apr-02 Release date: 01-May-02

Protein chain

No UniProt id for this chain

Struc: 155 a.a.

DOI no: 10.1074/jbc.M206342200

J Biol Chem 277:47476-47485 (2002)

PubMed id: 12207020

Crystal structure of OxyB, a cytochrome P450 implicated in an oxidative phenol coupling reaction during vancomycin biosynthesis.

K.Zerbe, O.Pylypenko, F.Vitali, W.Zhang, S.Rouset, M.Heck, J.W.Vrijbloed, D.Bischoff, B.Bister, R.D.Süssmuth, S.Pelzer, W.Wohlleben, J.A.Robinson, I.Schlichting.

ABSTRACT

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.

Selected figure(s)

Figure 1.
Fig. 1. Structures of glycopeptide antibiotics and putative linear heptapeptide intermediates (18-20) (1) in vancomycin biosynthesis.

Figure 2.
Fig. 2. The organization of ORFs in the DNA isolated from the vancomycin producer A. orientalis. On the left side is the 3'-terminal end of the peptide synthetase-3 and on the right side the 5'-terminal end of the halogenase gene (14). The oxyA, oxyB, and oxyC genes are indicated.

The above figures are reprinted by permission from the ASBMB: J Biol Chem (2002, 277, 47476-47485) copyright 2002.

Figures were selected by the author.

Author's comment

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 1.7Å 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 Cys347 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.
Ilme Schlichting