PDBsum entry 1mgo

Oxidoreductase

PDB id: 1mgo

Contents

Protein chains

374 a.a. *

Ligands

NAD ×2

PFB ×2

MPD

Metals

_ZN ×4

Waters ×465

* Residue conservation analysis

PDB id:

1mgo

Name:

Oxidoreductase

Title:

Horse liver alcohol dehydrogenase phe93ala mutant

Structure:

Alcohol dehydrogenase e chain. Chain: a, b. Engineered: yes. Mutation: yes

Source:

Equus caballus. Horse. Organism_taxid: 9796. Gene: m64864. Expressed in: escherichia coli. Expression_system_taxid: 562.

Biol. unit:

Dimer (from PQS)

Resolution:

1.20Å R-factor: 0.188 R-free: 0.209

Authors:

J.K.Rubach,B.V.Plapp

Key ref:

J.K.Rubach and B.V.Plapp (2002). Mobility of fluorobenzyl alcohols bound to liver alcohol dehydrogenases as determined by NMR and X-ray crystallographic studies. Biochemistry, 41, 15770-15779. PubMed id: 12501206 DOI: 10.1021/bi026581h

Date:

15-Aug-02 Release date: 13-Nov-02

Protein chains

P00327  (ADH1E_HORSE) -  Alcohol dehydrogenase E chain from Equus caballus

Seq: 375 a.a.

Struc: 374 a.a.*

* PDB and UniProt seqs differ at 1 residue position (black cross)

Enzyme reactions

• Enzyme class: E.C.1.1.1.1 - alcohol dehydrogenase.
• Reaction:
  1. a primary alcohol + NAD⁺ = an aldehyde + NADH + H⁺
  2. a secondary alcohol + NAD⁺ = a ketone + NADH + H⁺

primary alcohol + NAD(+) (Bound ligand (Het Group name = NAD) corresponds exactly) = aldehyde + NADH + H(+)

secondary alcohol + NAD(+) (Bound ligand (Het Group name = NAD) corresponds exactly) = ketone + NADH + H(+)

• Cofactor: Zn(2+) or Fe cation

Molecule diagrams generated from .mol files obtained from the KEGG ftp site

reference

DOI no: 10.1021/bi026581h

Biochemistry 41:15770-15779 (2002)

PubMed id: 12501206

Mobility of fluorobenzyl alcohols bound to liver alcohol dehydrogenases as determined by NMR and X-ray crystallographic studies.

J.K.Rubach, B.V.Plapp.

ABSTRACT

The relationship between substrate mobility and catalysis was studied with wild-type and Phe93Ala (F93A) horse liver alcohol dehydrogenase (ADH). Wild-type ADH binds 2,3,4,5,6-pentafluorobenzyl alcohol in one position as shown by X-ray results, and (19)F NMR shows five resonances for the fluorines of the bound alcohol. The two meta-fluorines exchange positions with a rate constant of about 4 s(-1), indicating that mobility (ring flipping) of the benzyl alcohol is relatively restricted. The wild-type enzyme binds 2,3-difluorobenzyl alcohol in two alternative conformations that are related by a ring flip and a small translation of the fluorinated benzene ring, and the (19)F NMR spectrum shows three resonances for the two bound fluorines, consistent with the two orientations. Phe-93 interacts with the bound benzyl alcohols, and the F93A substitution decreases the rate constants for hydride transfer for benzyl alcohol oxidation and benzaldehyde reduction by 7.4- and 130-fold, respectively. The structure of F93A ADH crystallized with NAD(+) and 2,3,4,5,6-pentafluorobenzyl alcohol is similar to the structure of the wild-type enzyme complex except that the pentafluorobenzyl alcohol is not found in one position. The (19)F NMR spectrum of the F93A ADH-NAD(+)-pentafluorobenzyl alcohol complex shows three resonances for the bound fluorines. Line shape analysis of the spectrum suggests the bound pentafluorobenzyl ring undergoes rapid ring-flipping at about 20 000 s(-1). The F93A substitution greatly increases the mobility of the benzyl alcohol but modestly and differentially decreases the probability that the substrate is preorganized for hydride transfer.