PDBsum entry: 2z39

Hydrolase

PDB-id: 2z39

Contents

Description

Header details

Header records

References

PROCHECK

Protein chains

246 a.a. *

Metal ions

_CL ×19

Waters ×421

* Residue conservation analysis

Tools

Clefts Calculation

PDB id:

2z39

Name:

Hydrolase

Title:

Crystal structure of brassica juncea chitinase catalytic module glu234ala mutant (bjchi3-e234a)

Structure:

Chitinase. Chain: a, b. Fragment: catalytic module, unp residues 146-389. Engineered: yes. Mutation: yes

Source:

Brassica juncea. Organism_taxid: 3707. Gene: bjchi1. Expressed in: pichia pastoris. Expression_system_taxid: 4922.

UniProt:

Chains A, B: Q9SQF7 (Q9SQF7_BRAJU)

Seq:

Struc:

Seq:

400 a.a.

Struc:

246 a.a.*

Key:	PfamA domain
Secondary structure	CATH domain

* PDB and UniProt seqs differ at 2 residue positions (black crosses)

Resolution:

1.70Å

R-factor:

0.180

R-free:

0.222

Authors:

W.Ubhayasekera,T.Bergfors,S.L.Mowbray

Key ref:

W.Ubhayasekera et al. (2007). Crystal structures of a family 19 chitinase from Brassica juncea show flexibility of binding cleft loops.. FEBS J, 274, 3695-3703. [PubMed id: 17608716] [DOI: 10.1111/j.1742-4658.2007.05906.x]

Date:

02-Jun-07

Release date:

19-Jun-07

Related entries:

2z37
brassica juncea chitinase catalytic module
2z38
brassica juncea chitinase catalytic module with bound
chloride ions

Quick_links

Procheck

Clefts

Surface

Key reference

DOI no: 10.1111/j.1742-4658.2007.05906.x

FEBS J 274:3695-3703 (2007)

PubMed id: 17608716

Crystal structures of a family 19 chitinase from Brassica juncea show flexibility of binding cleft loops.

W.Ubhayasekera, C.M.Tang, S.W.Ho, G.Berglund, T.Bergfors, M.L.Chye, S.L.Mowbray.

ABSTRACT

Brassica juncea chitinase is an endo-acting, pathogenesis-related protein that is classified into glycoside hydrolase family 19, with highest homology (50-60%) in its catalytic domain to class I plant chitinases. Here we report X-ray structures of the chitinase catalytic domain from wild-type (apo, as well as with chloride ions bound) and a Glu234Ala mutant enzyme, solved by molecular replacement and refined at 1.53, 1.8 and 1.7 A resolution, respectively. Confirming our earlier mutagenesis studies, the active-site residues are identified as Glu212 and Glu234. Glu212 is believed to be the catalytic acid in the reaction, whereas Glu234 is thought to have a dual role, both activating a water molecule in its attack on the anomeric carbon, and stabilizing the charged intermediate. The molecules in the various structures differ significantly in the conformation of a number of loops that border the active-site cleft. The differences suggest an opening and closing of the enzyme during the catalytic cycle. Chitin is expected to dock first near Glu212, which will protonate it. Conformational changes then bring Glu234 closer, allowing it to assist in the following steps. These observations provide important insights into catalysis in family 19 chitinases.