PDBsum entry: 2oxb

Hydrolase

PDB id: 2oxb

Contents

Protein chain

537 a.a. *

Ligands

NAG ×3

SUC

Waters ×34

* Residue conservation analysis

PDB id:

2oxb

Name:

Hydrolase

Title:

Crystal structure of a cell-wall invertase (e203q) from arab thaliana in complex with sucrose

Structure:

Beta-fructofuranosidase. Chain: a. Synonym: putative beta- fructofuranosidase 1. Engineered: yes. Mutation: yes

Source:

Arabidopsis thaliana. Thale cress. Organism_taxid: 3702. Gene: atbfruct1. Expressed in: pichia pastoris. Expression_system_taxid: 4922

Resolution:

2.60Å R-factor: 0.205 R-free: 0.268

Authors:

W.Lammens,K.Le Roy,A.Van Laere,W.Van Den Ende,A.Rabijns

Key ref:

J.Mátrai et al. (2008). An alternate sucrose binding mode in the E203Q Arabidopsis invertase mutant: an X-ray crystallography and docking study. Proteins, 71, 552-564. PubMed id: 17963237 DOI: 10.1002/prot.21700

Date:

20-Feb-07 Release date: 22-Jan-08

Protein chain

Q43866  (INV1_ARATH) -  Beta-fructofuranosidase, insoluble isoenzyme CWINV1

Seq: 584 a.a.

Struc: 537 a.a.*

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

Enzyme reactions

• Enzyme class: E.C.3.2.1.26 - Beta-fructofuranosidase.
• Reaction: Hydrolysis of terminal non-reducing beta-D-fructofuranoside residues in beta-D-fructofuranosides.

 Gene Ontology (GO) functional annotation 

• Cellular component: extracellular region (3 terms)
• Biological process: metabolic process (5 terms)
• Biochemical function: hydrolase activity (6 terms)

DOI no: 10.1002/prot.21700

Proteins 71:552-564 (2008)

PubMed id: 17963237

An alternate sucrose binding mode in the E203Q Arabidopsis invertase mutant: an X-ray crystallography and docking study.

J.Mátrai, W.Lammens, A.Jonckheer, K.Le Roy, A.Rabijns, W.Van den Ende, M.De Maeyer.

ABSTRACT

In the present study, we report on the X-ray crystallographic structure of a GH32 invertase mutant, (i.e., the Arabidopsis thaliana cell-wall invertase 1-E203Q, AtcwINV1-mutant) in complex with sucrose. This structure was solved to reveal the features of sugar binding in the catalytic pocket. However, as demonstrated by the X-ray structure the sugar binding and the catalytic pocket arrangement is significantly altered as compared with what was expected based on previous X-ray structures on GH-J clan enzymes. We performed a series of docking and molecular dynamics simulations on various derivatives of AtcwINV1 to reveal the reasons behind this modified sugar binding. Our results demonstrate that the E203Q mutation introduced into the catalytic pocket triggers conformational changes that alter the wild type substrate binding. In addition, this study also reveals the putative productive sucrose binding modus in the wild type enzyme.

Selected figure(s)

Figure 1.
Figure 1. (a) AtcwINV1 and (b) AtcwINV1-mutant-sucrose structures. The N-acetylglucosamine residues and glycosyltation sites are indicated by orange color. (c) and (d) panels show a zoom in into the catalytic pocket of the AtcwINV1 and AtcwINV1-mutant enzymes, respectively.

Figure 2.
Figure 2. Representation of the sucroses/raffinose as they appear in the superimposed X-ray structures. Color codes are as follows: green: FEH IIa/sucrose (the FM modus), red: levansucrase (E342A)/sucrose (the LSM modus), orange: -fructosidase/raffinose, blue: invertase(E203Q)/sucrose (the MIM modus).

The above figures are reprinted by permission from John Wiley & Sons, Inc.: Proteins (2008, 71, 552-564) copyright 2008.

Figures were selected by an automated process.