PDBsum entry 2nsf

Isomerase

PDB id: 2nsf

Contents

Protein chain

240 a.a. *

Ligands

SO4 ×3

GOL

Metals

_ZN

Waters ×229

* Residue conservation analysis

PDB id:

2nsf

Name:

Isomerase

Title:

Crystal structure of the mycothiol-dependent maleylpyruvate isomerase

Structure:

Hypothetical protein cgl3021. Chain: a. Synonym: maleylpyruvate isomerase. Engineered: yes

Source:

Corynebacterium glutamicum. Organism_taxid: 1718. Expressed in: escherichia coli bl21(de3). Expression_system_taxid: 469008.

Resolution:

1.75Å R-factor: 0.230 R-free: 0.234

Authors:

W.R.Chang,R.Wang

Key ref:

R.Wang et al. (2007). Crystal Structures and Site-directed Mutagenesis of a Mycothiol-dependent Enzyme Reveal a Novel Folding and Molecular Basis for Mycothiol-mediated Maleylpyruvate Isomerization. J Biol Chem, 282, 16288-15294. PubMed id: 17428791 DOI: 10.1074/jbc.M610347200

Date:

04-Nov-06 Release date: 10-Apr-07

Protein chain

Q8NLC1  (Q8NLC1_CORGL) -  Uncharacterized protein from Corynebacterium glutamicum (strain ATCC 13032 / DSM 20300 / JCM 1318 / BCRC 11384 / CCUG 27702 / LMG 3730 / NBRC 12168 / NCIMB 10025 / NRRL B-2784 / 534)

Seq: 241 a.a.

Struc: 240 a.a.

Enzyme reactions

• Enzyme class: E.C.5.2.1.4 - maleylpyruvate isomerase.
• Reaction: 3-maleylpyruvate = 3-fumarylpyruvate

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

Added reference

DOI no: 10.1074/jbc.M610347200

J Biol Chem 282:16288-15294 (2007)

PubMed id: 17428791

Crystal Structures and Site-directed Mutagenesis of a Mycothiol-dependent Enzyme Reveal a Novel Folding and Molecular Basis for Mycothiol-mediated Maleylpyruvate Isomerization.

R.Wang, Y.J.Yin, F.Wang, M.Li, J.Feng, H.M.Zhang, J.P.Zhang, S.J.Liu, W.R.Chang.

ABSTRACT

Mycothiol (MSH) is the major low molecular mass thiols in many Gram-positive bacteria such as Mycobacterium tuberculosis and Corynebacterium glutamicum. The physiological roles of MSH are believed to be equivalent to those of GSH in Gram-negative bacteria, but current knowledge of MSH is limited to detoxification of alkalating chemicals and protection from host cell defense/killing systems. Recently, an MSH-dependent maleylpyruvate isomerase (MDMPI) was discovered from C. glutamicum, and this isomerase represents one example of many putative MSH-dependent enzymes that take MSH as cofactor. In this report, fourteen mutants of MDMPI were generated. The wild type and mutant (H52A) MDMPIs were crystallized and their structures were solved at 1.75 and 2.05A resolution, respectively. The crystal structures reveal that this enzyme contains a divalent metal-binding domain and a C-terminal domain possessing a novel folding pattern (alphabetaalphabetabetaalpha fold). The divalent metal-binding site is composed of residues His(52), Glu(144), and His(148) and is located at the bottom of a surface pocket. Combining the structural and site-directed mutagenesis studies, it is proposed that this surface pocket including the metal ion and MSH moiety formed the putative catalytic center.

Selected figure(s)

Figure 4.
FIGURE 4. The side chain of Arg^222 (NH1) at the C-terminal domain interacts with Asp^151 (OD2) of the N-terminal domain through a salt bridge. The N-terminal domain is shown in yellow, C-terminal domain in cyan, and interdomain coil in gray. Asp^151 and Arg^222 are represented in stick model (carbon atoms are colored the same as the domain they locate at) and the metal ion by a sphere (gray). Salt bridge is shown by dashed line. The figure was created by PyMol.

Figure 6.
FIGURE 6. The stereo view of the conformation differences in the putative active pocket between the wild type and the mutant (H52A) MDMPI. Mutant H52A is shown in purple and native MDMPI in green. Conformation differences were found at the side chains of Arg^82 and Trp^44. In H52A the metal ion was missing and a glycerol occupied the position of the metal ion, and the location of SO^2–[4] also changed. Furthermore, there is no obvious conformation changes observed, when compared with that of the wild type. The image was created by PyMol.

The above figures are reprinted by permission from the ASBMB: J Biol Chem (2007, 282, 16288-15294) copyright 2007.

Figures were selected by an automated process.