PDBsum entry 2oui

Oxidoreductase

PDB id: 2oui

Contents

Protein chains

360 a.a. *

Ligands

NO3 ×4

CAC ×4

EDO ×9

PGE ×4

1PE

PG4

Metals

_ZN ×4

_CL

Waters ×1085

* Residue conservation analysis

PDB id:

2oui

Name:

Oxidoreductase

Title:

D275p mutant of alcohol dehydrogenase from protozoa entamoeba histolytica

Structure:

NADP-dependent alcohol dehydrogenase. Chain: a, b, c, d. Engineered: yes. Mutation: yes

Source:

Entamoeba histolytica. Organism_taxid: 5759. Gene: adh1. Expressed in: escherichia coli. Expression_system_taxid: 562.

Resolution:

1.77Å R-factor: 0.150 R-free: 0.178

Authors:

F.Frolow,L.Shimon,Y.Burstein,E.Goihberg,M.Peretz,O.Dym

Key ref:

E.Goihberg et al. (2008). Thermal stabilization of the protozoan Entamoeba histolytica alcohol dehydrogenase by a single proline substitution. Proteins, 72, 711-719. PubMed id: 18260103 DOI: 10.1002/prot.21946

Date:

11-Feb-07 Release date: 12-Feb-08

Protein chains

P35630  (ADH1_ENTHI) -  NADP-dependent isopropanol dehydrogenase from Entamoeba histolytica (strain ATCC 30459 / HM-1:IMSS / ABRM)

Seq: 366 a.a.

Struc: 360 a.a.*

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

Enzyme reactions

• Enzyme class: E.C.1.1.1.80 - isopropanol dehydrogenase (NADP(+)).
• Reaction: propan-2-ol + NADP⁺ = acetone + NADPH + H⁺

propan-2-ol + NADP(+) (Bound ligand (Het Group name = EDO) matches with 60.00% similarity) = acetone + NADPH + H(+)

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

reference

DOI no: 10.1002/prot.21946

Proteins 72:711-719 (2008)

PubMed id: 18260103

Thermal stabilization of the protozoan Entamoeba histolytica alcohol dehydrogenase by a single proline substitution.

E.Goihberg, O.Dym, S.Tel-Or, L.Shimon, F.Frolow, M.Peretz, Y.Burstein.

ABSTRACT

Analysis of the three-dimensional structures of two closely related thermophilic and hyperthermophilic alcohol dehydrogenases (ADHs) from the respective microorganisms Entamoeba histolytica (EhADH1) and Thermoanaerobacter brockii (TbADH) suggested that a unique, strategically located proline residue (Pro275) at the center of the dimerization interface might be crucial for maintaining the thermal stability of TbADH. To assess the contribution of Pro275 to the thermal stability of the ADHs, we applied site-directed mutagenesis to replace Asp275 of EhADH1 with Pro (D275P-EhADH1) and conversely Pro275 of TbADH with Asp (P275D-TbADH). The results indicate that replacing Asp275 with Pro significantly enhances the thermal stability of EhADH1 (DeltaT(1/2) <or= +10 degrees C), whereas the reverse mutation in the thermophilic TbADH (P275D-TbADH) reduces the thermostability of the enzyme (DeltaT(1/2) <or= -18.8 degrees C). Analysis of the crystal structures of the thermostabilized mutant D275P-EhADH1 and the thermocompromised mutant P275D-TbADH suggest that a proline residue at position 275 thermostabilized the enzymes by reducing flexibility and by reinforcing hydrophobic interactions at the dimer-dimer interface of the tetrameric ADHs.

Selected figure(s)

Figure 1.
Figure 1. Structure-based sequence alignment of ADHs from Thermoanaerobacter brockii (TbADH, TB[18]), Clostridium beijerinckii (CbADH, CB[18]) and Entamoeba histolytica (EhADH, EH[19]). The secondary structure elements are shown above the sequences with twisted rods for -helices and arrows for -strands. The mutagenesis site (m) and the metal ligand residues (Zn) are depicted above and below the sequence, respectively. The figure was prepared with INDONESIA program package http://alpha2.bmc.uu.se/ [sim.gif]dermis/.

Figure 2.
Figure 2. (a) Ribbon diagram of the TbADH tetramer. Subunits are represented in different colors. Pro residues, depicted in ball representation, are colored magenta (Pro275) and cyan (Pro100). The active site Zn is colored green. (b) Superimposition of the structures of the wild-type apo-EhADH1 (colored orange) and the apo D275P-EhADH1 mutant (colored green). Residues within a sphere of a 4-Å radius of the mutation are shown (the superscript refers to the subunit). (c) Superimposition of the structures of the wild-type holo-TbADH (colored green) and the holo P275D-TbADH mutant (colored orange). Residues within a sphere of a 4-Å radius of the mutation are shown (the superscript refers to the subunit).

The above figures are reprinted by permission from John Wiley & Sons, Inc.: Proteins (2008, 72, 711-719) copyright 2008.

Figures were selected by an automated process.