PDBsum entry: 1flg

Oxidoreductase

PDB id: 1flg

Contents

Protein chains

582 a.a. *

Ligands

PQQ ×2

Metals

_CA ×4

Waters ×106

* Residue conservation analysis

PDB id:

1flg

Name:

Oxidoreductase

Title:

Crystal structure of the quinoprotein ethanol dehydrogenase from pseudomonas aeruginosa

Structure:

Protein (quinoprotein ethanol dehydrogenase). Chain: a, b

Source:

Pseudomonas aeruginosa. Organism_taxid: 287

Biol. unit:

Dimer (from PQS)

Resolution:

2.60Å R-factor: 0.142 R-free: 0.274

Authors:

T.Keitel,A.Diehl,T.Knaute,J.J.Stezowski,W.Hohne,H.Gorisch

Key ref:

T.Keitel et al. (2000). X-ray structure of the quinoprotein ethanol dehydrogenase from Pseudomonas aeruginosa: basis of substrate specificity. J Mol Biol, 297, 961-974. PubMed id: 10736230 DOI: 10.1006/jmbi.2000.3603

Date:

14-Aug-00 Release date: 30-Aug-00

Supersedes:

1eee

Protein chains

Q9Z4J7  (EXAA_PSEAE) -  Quinoprotein ethanol dehydrogenase

Seq: 623 a.a.

Struc: 582 a.a.

 Gene Ontology (GO) functional annotation 

• Cellular component: membrane (3 terms)
• Biological process: oxidation reduction (1 term)
• Biochemical function: oxidoreductase activity (4 terms)

DOI no: 10.1006/jmbi.2000.3603

J Mol Biol 297:961-974 (2000)

PubMed id: 10736230

X-ray structure of the quinoprotein ethanol dehydrogenase from Pseudomonas aeruginosa: basis of substrate specificity.

T.Keitel, A.Diehl, T.Knaute, J.J.Stezowski, W.Höhne, H.Görisch.

ABSTRACT

The homodimeric enzyme form of quinoprotein ethanol dehydrogenase from Pseudomonas aeruginosa ATCC 17933 crystallizes readily with the space group R3. The X-ray structure was solved at 2.6 A resolution by molecular replacement.Aside from differences in some loops, the folding of the enzyme is very similar to the large subunit of the quinoprotein methanol dehydrogenases from Methylobacterium extorquens or Methylophilus W3A1. Eight W-shaped beta-sheet motifs are arranged circularly in a propeller-like fashion forming a disk-shaped superbarrel. No electron density for a small subunit like that in methanol dehydrogenase could be found. The prosthetic group is located in the centre of the superbarrel and is coordinated to a calcium ion. Most amino acid residues found in close contact with the prosthetic group pyrroloquinoline quinone and the Ca(2+) are conserved between the quinoprotein ethanol dehydrogenase structure and that of the methanol dehydrogenases. The main differences in the active-site region are a bulky tryptophan residue in the active-site cavity of methanol dehydrogenase, which is replaced by a phenylalanine and a leucine side-chain in the ethanol dehydrogenase structure and a leucine residue right above the pyrrolquinoline quinone group in methanol dehydrogenase which is replaced by a tryptophan side-chain. Both amino acid exchanges appear to have an important influence, causing different substrate specificities of these otherwise very similar enzymes. In addition to the Ca(2+) in the active-site cavity found also in methanol dehydrogenase, ethanol dehydrogenase contains a second Ca(2+)-binding site at the N terminus, which contributes to the stability of the native enzyme.

Selected figure(s)

Figure 1.
Figure 1. General folding topology of QEDH from P. aeruginosa with PQQ in the active-site and the two calcium ions (green) at the PQQ binding site (S1) and at an N-terminal binding site (S2). The eight propeller blades W[1] to W[8] are formed by four antiparallel b-strands A, B, C and D. The A strands are the innermost ones.

Figure 5.
Figure 5. Active-site cavity of QEDH from P. aeruginosa, including PQQ and Ca^2+, stereo view from the entrance to the active-site. (a) Ball-and-stick representation; (b) space-filling representation. Hydrophobic residues, brown; polar residues, yellow; acidic residues, red; PQQ, green; Ca^2+, blue.

The above figures are reprinted by permission from Elsevier: J Mol Biol (2000, 297, 961-974) copyright 2000.

Figures were selected by an automated process.