PDBsum entry 1j6t

Transferase

PDB id: 1j6t

Contents

Protein chains

144 a.a. *

85 a.a. *

Ligands

PO3

* Residue conservation analysis

PDB id:

1j6t

Name:

Transferase

Title:

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

Structure:

Pts system, mannitol-specific iiabc component. Chain: a. Fragment: eiia domain. Synonym: iiamtl, eiia-mtl, phosphotransferase enzyme ii, a domain component. Engineered: yes. Phosphocarrier protein hpr. Chain: b. Synonym: hpr, histidine-containing protein.

Source:

Escherichia coli. Organism_taxid: 562. Expressed in: escherichia coli. Expression_system_taxid: 562. Expression_system_taxid: 562

NMR struc:

3 models

Authors:

G.M.Clore,G.Cornilescu

Key ref:

G.Cornilescu et al. (2002). Solution structure of the phosphoryl transfer complex between the cytoplasmic A domain of the mannitol transporter IIMannitol and HPr of the Escherichia coli phosphotransferase system. J Biol Chem, 277, 42289-42298. PubMed id: 12202490 DOI: 10.1074/jbc.M207314200

Date:

14-Aug-02 Release date: 13-Nov-02

Protein chain

P00550  (PTM3C_ECOLI) -  PTS system mannitol-specific EIICBA component from Escherichia coli (strain K12)

Seq: 637 a.a.

Struc: 144 a.a.

Protein chain

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

Seq: 85 a.a.

Struc: 85 a.a.

Enzyme reactions

• Enzyme class 2: Chain A: E.C.2.7.1.197 - protein-N(pi)-phosphohistidine--D-mannitol phosphotransferase.
• Reaction: D-mannitol(out) + N(pros)-phospho-L-histidyl-[protein] = D-mannitol 1-phosphate(in) + L-histidyl-[protein]
• Enzyme class 3: Chain B: E.C.?

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

Key reference

DOI no: 10.1074/jbc.M207314200

J Biol Chem 277:42289-42298 (2002)

PubMed id: 12202490

Solution structure of the phosphoryl transfer complex between the cytoplasmic A domain of the mannitol transporter IIMannitol and HPr of the Escherichia coli phosphotransferase system.

G.Cornilescu, B.R.Lee, C.C.Cornilescu, G.Wang, A.Peterkofsky, G.M.Clore.

ABSTRACT

The solution structure of the complex between the cytoplasmic A domain (IIA(Mtl)) of the mannitol transporter II(Mannitol) and the histidine-containing phosphocarrier protein (HPr) of the Escherichia coli phosphotransferase system has been solved by NMR, including the use of conjoined rigid body/torsion angle dynamics, and residual dipolar couplings, coupled with cross-validation, to permit accurate orientation of the two proteins. A convex surface on HPr, formed by helices 1 and 2, interacts with a complementary concave depression on the surface of IIA(Mtl) formed by helix 3, portions of helices 2 and 4, and beta-strands 2 and 3. The majority of intermolecular contacts are hydrophobic, with a small number of electrostatic interactions at the periphery of the interface. The active site histidines, His-15 of HPr and His-65 of IIA(Mtl), are in close spatial proximity, and a pentacoordinate phosphoryl transition state can be readily accommodated with no change in protein-protein orientation and only minimal perturbations of the backbone immediately adjacent to the histidines. Comparison with two previously solved structures of complexes of HPr with partner proteins of the phosphotransferase system, the N-terminal domain of enzyme I (EIN) and enzyme IIA(Glucose) (IIA(Glc)), reveals a number of common features despite the fact that EIN, IIA(Glc), and IIA(Mtl) bear no structural resemblance to one another. Thus, entirely different underlying structural elements can form binding surfaces for HPr that are similar in terms of both shape and residue composition. These structural comparisons illustrate the roles of surface and residue complementarity, redundancy, incremental build-up of specificity and conformational side chain plasticity in the formation of transient specific protein-protein complexes in signal transduction pathways.

Selected figure(s)

Figure 5.
Fig. 5. The transition state of the IIA^Mtl-P-HPr complex. A, detailed view around the active site histidines, illustrating the backbone and side chain positions in the unphosphorylated complex, the dissociative transition state, and the associative transition state. The backbones of IIA^Mtl and HPr are shown in dark blue and dark green, respectively, for the unphosphorylated complex, and in light blue and light green, respectively, for the putative dissociative and associative transition states; the active site histidines and pentacoordinate phosphoryl group (in the case of the transition states) are shown in purple for the unphosphorylated complex, in red for the dissociative transition state (N 2-N 1 distance of ~6 Å between His-65 and His-15), and in orange for the associative transition state (N 2-N 1 distance of ~4 Å between His-65 and His-15). Small changes in the backbone of residues 64-66 of IIA^Mtl and residues 14-16 of HPr are required to accommodate the transition states. B, detailed view of the active site in the putative transition state illustrating the interactions that stabilize the phosphoryl group. The color coding is as follows: the backbone and side chains of IIA^Mtl are shown in blue and red, respectively; the backbone and side chains of HPr are shown in green and gray, respectively; the active site histidines are in purple, and the pentacoordinate phosphoryl group is in yellow. Residues from HPr are labeled in italics.

Figure 6.
Fig. 6. Surface representations illustrating the binding surfaces involved in the IIA^Mtl-HPr (left panel), IIA^Glc-HPr (middle panel), and EIN-HPr (right panel) complexes. The binding surfaces on IIA^Mtl, IIA Glc, and EIN are shown in A, and the binding surfaces on HPr are shown in B. The binding surfaces are color-coded with hydrophobic residues in green, polar residues in light blue, the active site histidines in purple, positively charged residues in dark blue, and negatively charged residues in red. The relevant portions of the backbone of HPr are shown in gold in A; the relevant portions of the backbone of IIA^Mtl, IIA^Glc, and EIN are shown in blue, red, and green, respectively, in B. The location of the phosphoryl group in the transition state is depicted in yellow. Residues of HPr are labeled in italics. The coordinates of the EIN-HPr and IIA^Glc-HPr complexes are taken from Garrett et al. (16) (code 3EZE) and Wang et al. (17) (code 1GGR), respectively.

The above figures are reprinted by permission from the ASBMB: J Biol Chem (2002, 277, 42289-42298) copyright 2002.

Figures were selected by an automated process.