PDBsum entry 3l1v

Hydrolase

PDB id: 3l1v

Contents

Protein chains

182 a.a. *

Ligands

PO4 ×2

Metals

_ZN ×2

_CA ×2

Waters ×251

* Residue conservation analysis

PDB id:

3l1v

Name:

Hydrolase

Title:

Crystal structure of gmhb from e. Coli in complex with calcium and phosphate.

Structure:

D,d-heptose 1,7-bisphosphate phosphatase. Chain: a, b. Synonym: d-glycero-d-manno-heptose 1,7-bisphosphate phosphatase. Engineered: yes

Source:

Escherichia coli. Organism_taxid: 83333. Strain: k-12. Gene: b0200, gmhb, jw0196, yaed. Expressed in: escherichia coli. Expression_system_taxid: 562.

Resolution:

1.95Å R-factor: 0.249 R-free: 0.284

Authors:

S.N.Sugiman-Marangos,M.S.Junop

Key ref:

P.L.Taylor et al. (2010). Structural and kinetic characterization of the LPS biosynthetic enzyme D-alpha,beta-D-heptose-1,7-bisphosphate phosphatase (GmhB) from Escherichia coli. Biochemistry, 49, 1033-1041. PubMed id: 20050699

Date:

14-Dec-09 Release date: 05-Jan-10

Protein chains

P63228  (GMHBB_ECOLI) -  D-glycero-beta-D-manno-heptose-1,7-bisphosphate 7-phosphatase from Escherichia coli (strain K12)

Seq: 191 a.a.

Struc: 182 a.a.

Enzyme reactions

• Enzyme class: E.C.3.1.3.82 - D-glycero-beta-D-manno-heptose 1,7-bisphosphate 7-phosphatase.
• Reaction: D-glycero-beta-D-manno-heptose 1,7-bisphosphate + H2O = D-glycero-beta-D- manno-heptose 1-phosphate + phosphate

D-glycero-beta-D-manno-heptose 1,7-bisphosphate + H2O = D-glycero-beta-D- manno-heptose 1-phosphate + phosphate (Bound ligand (Het Group name = PO4) corresponds exactly)

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

Key reference

Biochemistry 49:1033-1041 (2010)

PubMed id: 20050699

Structural and kinetic characterization of the LPS biosynthetic enzyme D-alpha,beta-D-heptose-1,7-bisphosphate phosphatase (GmhB) from Escherichia coli.

P.L.Taylor, S.Sugiman-Marangos, K.Zhang, M.A.Valvano, G.D.Wright, M.S.Junop.

ABSTRACT

Lipopolysaccharide is a major component of the outer membrane of gram-negative bacteria and provides a permeability barrier to many commonly used antibiotics. ADP-heptose residues are an integral part of the LPS inner core, and mutants deficient in heptose biosynthesis demonstrate increased membrane permeability. The heptose biosynthesis pathway involves phosphorylation and dephosphorylation steps not found in other pathways for the synthesis of nucleotide sugar precursors. Consequently, the heptose biosynthetic pathway has been marked as a novel target for antibiotic adjuvants, which are compounds that facilitate and potentiate antibiotic activity. D-alpha,beta-D-heptose-1,7-bisphosphate phosphatase (GmhB) catalyzes the third essential step of LPS heptose biosynthesis. This study describes the first crystal structure of GmhB and enzymatic analysis of the protein. Structure-guided mutations followed by steady state kinetic analysis, together with established precedent for HAD phosphatases, suggest that GmhB functions through a phosphoaspartate intermediate. This study provides insight into the structure-function relationship of GmhB, a new target for combatting gram-negative bacterial infection.