PDBsum entry 2bv4

Lectin

PDB id: 2bv4

Contents

Protein chains

113 a.a. *

Ligands

MMA ×2

Metals

_CA ×4

Waters ×380

* Residue conservation analysis

PDB id:

2bv4

Name:

Lectin

Title:

1.0a structure of chromobacterium violaceum lectin in complex with alpha-methyl-mannoside

Structure:

Lectin cv-iil. Chain: a, b. Engineered: yes

Source:

Chromobacterium violaceum. Organism_taxid: 536. Expressed in: escherichia coli. Expression_system_taxid: 562.

Biol. unit:

Tetramer (from PDB file)

Resolution:

1.00Å R-factor: 0.107 R-free: 0.123

Authors:

M.Pokorna,G.Cioci,S.Perret,E.Rebuffet,J.Adam,N.Gilboa-Garber, E.P.Mitchell,A.Imberty,M.Wimmerova

Key ref:

M.Pokorná et al. (2006). Unusual entropy-driven affinity of Chromobacterium violaceum lectin CV-IIL toward fucose and mannose. Biochemistry, 45, 7501-7510. PubMed id: 16768446 DOI: 10.1021/bi060214e

Date:

22-Jun-05 Release date: 25-May-06

Protein chains

Q7NX84  (Q7NX84_CHRVO) -  Calcium-mediated lectin domain-containing protein from Chromobacterium violaceum (strain ATCC 12472 / DSM 30191 / JCM 1249 / NBRC 12614 / NCIMB 9131 / NCTC 9757)

Seq: 114 a.a.

Struc: 113 a.a.

DOI no: 10.1021/bi060214e

Biochemistry 45:7501-7510 (2006)

PubMed id: 16768446

Unusual entropy-driven affinity of Chromobacterium violaceum lectin CV-IIL toward fucose and mannose.

M.Pokorná, G.Cioci, S.Perret, E.Rebuffet, N.Kostlánová, J.Adam, N.Gilboa-Garber, E.P.Mitchell, A.Imberty, M.Wimmerová.

ABSTRACT

The purple pigmented bacterium Chromobacterium violaceum is a dominant component of tropical soil microbiota that can cause rare but fatal septicaemia in humans. Its sequenced genome provides insight into the abundant potential of this organism for biotechnological and pharmaceutical applications and allowed an ORF encoding a protein that is 60% identical to the fucose binding lectin (PA-IIL) from Pseudomonas aeruginosa and the mannose binding lectin (RS-IIL) from Ralstonia solanacearum to be identified. The lectin, CV-IIL, has recently been purified from C. violaceum [Zinger-Yosovich, K., Sudakevitz, D., Imberty, A., Garber, N. C., and Gilboa-Garber, N. (2006) Microbiology 152, 457-463] and has been confirmed to be a tetramer with subunit size of 11.86 kDa and a binding preference for fucose. We describe here the cloning of CV-IIL and its expression as a recombinant protein. A complete structure-function characterization has been made in an effort to analyze the specificity and affinity of CV-IIL for fucose and mannose. Crystal structures of CV-IIL complexes with monosaccharides have yielded the molecular basis of the specificity. Each monomer contains two close calcium cations that mediate the binding of the monosaccharides, which occurs in different orientations for fucose and mannose. The thermodynamics of binding has been analyzed by titration microcalorimetry, giving dissociation constants of 1.7 and 19 microM for alpha-methyl fucoside and alpha-methyl mannoside, respectively. Further analysis demonstrated a strongly favorable entropy term that is unusual in carbohydrate binding. A comparison with both PA-IIL and RS-IIL, which have binding preferences for fucose and mannose, respectively, yielded insights into the monosaccharide specificity of this important class of soluble bacterial lectins.