PDBsum entry 2v0h

Transferase

PDB id: 2v0h

Contents

Protein chain

450 a.a. *

Ligands

PEG ×4

SO4 ×5

Metals

_CO ×2

Waters ×497

* Residue conservation analysis

PDB id:

2v0h

Name:

Transferase

Title:

Characterization of substrate binding and catalysis of the potential antibacterial target n-acetylglucosamine-1-phosphate uridyltransferase (glmu)

Structure:

Bifunctional protein glmu. Chain: a. Synonym: glmu from haemophilis influenzae. Ec: 2.-.-.-

Source:

Haemophilus influenzae. Organism_taxid: 727

Resolution:

1.79Å R-factor: 0.192 R-free: 0.214

Authors:

I.Mochalkin,S.Lightle,J.F.Ohren,N.Y.Chirgadze

Key ref:

I.Mochalkin et al. (2007). Characterization of substrate binding and catalysis in the potential antibacterial target N-acetylglucosamine-1-phosphate uridyltransferase (GlmU). Protein Sci, 16, 2657-2666. PubMed id: 18029420 DOI: 10.1110/ps.073135107

Date:

14-May-07 Release date: 15-Jan-08

Protein chain

P43889  (GLMU_HAEIN) -  Bifunctional protein GlmU from Haemophilus influenzae (strain ATCC 51907 / DSM 11121 / KW20 / Rd)

Seq: 456 a.a.

Struc: 450 a.a.

Enzyme reactions

• Enzyme class 1: E.C.2.3.1.157 - glucosamine-1-phosphate N-acetyltransferase.
• Pathway: UDP-N-acetylglucosamine Biosynthesis
• Reaction: alpha-D-glucosamine 1-phosphate + acetyl-CoA = N-acetyl-alpha-D- glucosamine 1-phosphate + CoA + H⁺
• Enzyme class 2: E.C.2.7.7.23 - UDP-N-acetylglucosamine diphosphorylase.

Pathway:

• Reaction: N-acetyl-alpha-D-glucosamine 1-phosphate + UTP + H⁺ = UDP-N-acetyl- alpha-D-glucosamine + diphosphate

Note, where more than one E.C. class is given (as above), each may correspond to a different protein domain or, in the case of polyprotein precursors, to a different mature protein.

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

reference

DOI no: 10.1110/ps.073135107

Protein Sci 16:2657-2666 (2007)

PubMed id: 18029420

Characterization of substrate binding and catalysis in the potential antibacterial target N-acetylglucosamine-1-phosphate uridyltransferase (GlmU).

I.Mochalkin, S.Lightle, Y.Zhu, J.F.Ohren, C.Spessard, N.Y.Chirgadze, C.Banotai, M.Melnick, L.McDowell.

ABSTRACT

N-Acetylglucosamine-1-phosphate uridyltransferase (GlmU) catalyzes the first step in peptidoglycan biosynthesis in both Gram-positive and Gram-negative bacteria. The products of the GlmU reaction are essential for bacterial survival, making this enzyme an attractive target for antibiotic drug discovery. A series of Haemophilus influenzae GlmU (hiGlmU) structures were determined by X-ray crystallography in order to provide structural and functional insights into GlmU activity and inhibition. The information derived from these structures was combined with biochemical characterization of the K25A, Q76A, D105A, Y103A, V223A, and E224A hiGlmU mutants in order to map these active-site residues to catalytic activity of the enzyme and refine the mechanistic model of the GlmU uridyltransferase reaction. These studies suggest that GlmU activity follows a sequential substrate-binding order that begins with UTP binding noncovalently to the GlmU enzyme. The uridyltransferase active site then remains in an open apo-like conformation until N-acetylglucosamine-1-phosphate (GlcNAc-1-P) binds and induces a conformational change at the GlcNAc-binding subsite. Following the binding of GlcNAc-1-P to the UTP-charged uridyltransferase active site, the non-esterified oxygen of GlcNAc-1-P performs a nucleophilic attack on the alpha-phosphate group of UTP. The new data strongly suggest that the mechanism of phosphotransfer in the uridyltransferase reaction in GlmU is primarily through an associative mechanism with a pentavalent phosphate intermediate and an inversion of stereochemistry. Finally, the structural and biochemical characterization of the uridyltransferase active site and catalytic mechanism described herein provides a basis for the structure-guided design of novel antibacterial agents targeting GlmU activity.

Selected figure(s)

Figure 2.
Figure 2. hiGlmU uridyltransferase active site. (A) Stereoview of the (Fo-Fc) OMIT electron density maps of UDP-GlcNAc, UDP, and uridine bound to the

Figure 4.
Figure 4. Structural insights into the mechanism of uridylation. (A) View of the GlmU uridyltransferase active site (open conformation) in the UDP-bound

The above figures are reprinted by permission from the Protein Society: Protein Sci (2007, 16, 2657-2666) copyright 2007.

Figures were selected by an automated process.