PDBsum entry 1vrc

Transferase

PDB id: 1vrc

Contents

Protein chains

129 a.a. *

85 a.a. *

Ligands

PO3 ×2

* Residue conservation analysis

PDB id:

1vrc

Name:

Transferase

Title:

Complex of enzyme iiamannose and the histidine-containing phosphocarrier protein hpr from escherichia coli nmr, restrained regularized mean structure

Structure:

Pts system, mannose-specific iiab component. Chain: a, b. Fragment: eiia domain. Synonym: eiiab-man, mannose-permease iiab component, phosphotransferase enzyme ii, ab component, eiii-man. Engineered: yes. Phosphocarrier protein hpr. Chain: c, d. Synonym: histidine-containing protein.

Source:

Escherichia coli. Organism_taxid: 562. Gene: manx, gptb, ptsl. Expressed in: escherichia coli. Expression_system_taxid: 562. Gene: ptsh, hpr. Expression_system_taxid: 562

NMR struc:

2 models

Authors:

G.M.Clore,D.C.Williams

Key ref:

D.C.Williams et al. (2005). Solution NMR structure of the 48-kDa IIAMannose-HPr complex of the Escherichia coli mannose phosphotransferase system. J Biol Chem, 280, 20775-20784. PubMed id: 15788390 DOI: 10.1074/jbc.M501986200

Date:

21-Feb-05 Release date: 19-Apr-05

Protein chains

P69797  (PTNAB_ECOLI) -  PTS system mannose-specific EIIAB component from Escherichia coli (strain K12)

Seq: 323 a.a.

Struc: 129 a.a.

Protein chains

P0AA04  (PTHP_ECOLI) -  Phosphocarrier protein HPr from Escherichia coli (strain K12)

Seq: 85 a.a.

Struc: 85 a.a.

Enzyme reactions

• Enzyme class: Chains A, B: E.C.2.7.1.191 - protein-N(pi)-phosphohistidine--D-mannose phosphotransferase.
• Reaction: D-mannose(out) + N(pros)-phospho-L-histidyl-[protein] = D-mannose 6-phosphate(in) + L-histidyl-[protein]

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

Key reference

DOI no: 10.1074/jbc.M501986200

J Biol Chem 280:20775-20784 (2005)

PubMed id: 15788390

Solution NMR structure of the 48-kDa IIAMannose-HPr complex of the Escherichia coli mannose phosphotransferase system.

D.C.Williams, M.Cai, J.Y.Suh, A.Peterkofsky, G.M.Clore.

ABSTRACT

The solution structure of the 48-kDa IIA(Man)-HPr complex of the mannose branch of the Escherichia coli phosphotransferase system has been solved by NMR using conjoined rigid body/torsion angle-simulated annealing on the basis of intermolecular nuclear Overhauser enhancement data and residual dipolar couplings. IIA(Man) is dimeric and has two symmetrically related binding sites per dimer for HPr. A convex surface on HPr, formed primarily by helices 1 and 2, interacts with a deep groove at the interface of the two subunits of IIA(Man). The interaction surface on IIA(Man) is predominantly helical, comprising helix 3 from the subunit that bears the active site His-10 and helices 1, 4, and 5 from the other subunit. The total buried accessible surface area at the protein-protein interface is 1450 A(2). The binding sites on the two proteins are complementary in terms of shape and distribution of hydrophobic, hydrophilic, and charged residues. The active site histidines, His-10 of IIA(Man) and His-15 (italics indicate HPr residues) of HPr, are in close proximity. An associative transition state involving a pentacoordinate phosphoryl group with trigonal bipyramidal geometry bonded to the N-epsilon2 atom of His-10 and the N-delta1 atom of His-15 can be readily formed with negligible displacement in the backbone coordinates of the residues immediately adjacent to the active site histidines. Comparing the structures of complexes of HPr with three other structurally unrelated phosphotransferase system proteins, enzymes I, IIA(glucose), and IIA(mannitol), reveals a number of common features that provide a molecular basis for understanding how HPr specifically recognizes a wide range of diverse proteins.

Selected figure(s)

Figure 5.
FIG. 5. The IIA^Man-HPr interface. A, stereoview of the interface with the backbone shown as tubes (green, HPr; blue, subunit A of IIA^Man; red, subunit B of IIA^Man) and the side chains as bonds (light green, HPr; cyan, subunit A of IIA^Man; pink, subunit B of IIA^Man; purple, active site histidines). Residues of HPr are labeled in italics. Note that the 1-ppm chemical shift perturbation observed for the backbone amide of Ser-72 of IIA^Man (cf. Fig. 1) is because of a ring current shift arising from its close proximity ( 3.9 Å) to the imidazole ring of His-15 of HPr. No other backbone amide proton of IIA^Man is within 5.7 Å of an aromatic side chain of HPr. B, diagrammatic summary of interfacial contacts. Residuesinvolved in potential electrostatic, hydrogen-bonding, or water-mediated hydrogen-bonding interactions are indicated in red (side chain-side chain contacts), blue (side chain of HPr to backbone carbonyl of IIA^Man), or green (side chain of IIA^Man to backbone carbonyl of HPr). The active site histidines are shown in purple. C, interaction surface on IIA^Man for binding HPr. D, interaction surface on HPr for binding IIA^Man. Residues in the interaction surfaces (C and D) are color-coded as hydrophobic (green), hydrophilic (cyan), positively charged (blue), or negatively charged (red). The relevant portions of the backbone of the interacting partner, HPr in C (gold) and IIA^Man in D (lilac for subunit A, gold for subunit B,) are displayed as tubes. In C, the surface of the non-interacting residues of IIA^Man is colored in dark gray for subunit A and light gray for subunit B. Residues of HPr are labeled in italics.

Figure 6.
FIG. 6. Comparison of the relative orientation of the interacting helices in the IIA^Man-HPr and EIN-HPr complexes. The two complexes have been best-fitted to the coordinates of HPr. Two approximately orthogonal views are shown with the helices of HPr ( 1 and 2) in green; helices 1, 4, and 5 of subunit B of IIA^Man in red; helix 3 of subunit A of IIA^Man in blue; and helices 2, 2', 3, and 4 of EIN in gold. The coordinates of the EIN-HPr complex are taken from Ref. 5 (Protein Data Bank accession code 3EZA [PDB] ).

The above figures are reprinted from an Open Access publication published by the ASBMB: J Biol Chem (2005, 280, 20775-20784) copyright 2005.

Figures were selected by an automated process.