PDBsum entry 2fmi

Signaling protein

PDB id: 2fmi

Contents

Protein chains

128 a.a. *

14 a.a. *

Ligands

SO4

TRS

GOL

Waters ×87

* Residue conservation analysis

PDB id:

2fmi

Name:

Signaling protein

Title:

Crystal structure of chey in complex with chez 200-214 solved from a f432 crystal grown in tris (ph 8.4)

Structure:

Chemotaxis protein chey. Chain: a. Engineered: yes. C-terminal 15-mer from chemotaxis protein chez. Chain: b. Fragment: residues 200-214. Engineered: yes

Source:

Salmonella typhimurium. Organism_taxid: 99287. Strain: lt2. Gene: chey. Expressed in: escherichia coli. Expression_system_taxid: 562. Synthetic: yes. Other_details: this sequence corresponds to thE C-terminal 15 residues of the chez protein occurring naturally in salmonella

Biol. unit:

Dimer (from PQS)

Resolution:

2.30Å R-factor: 0.196 R-free: 0.219

Authors:

J.Guhaniyogi,V.L.Robinson,A.M.Stock

Key ref:

J.Guhaniyogi et al. (2006). Crystal structures of beryllium fluoride-free and beryllium fluoride-bound CheY in complex with the conserved C-terminal peptide of CheZ reveal dual binding modes specific to CheY conformation. J Mol Biol, 359, 624-645. PubMed id: 16674976 DOI: 10.1016/j.jmb.2006.03.050

Date:

09-Jan-06 Release date: 23-May-06

Protein chain

P0A2D5  (CHEY_SALTY) -  Chemotaxis protein CheY from Salmonella typhimurium (strain LT2 / SGSC1412 / ATCC 700720)

Seq: 129 a.a.

Struc: 128 a.a.

Protein chain

P07800  (CHEZ_SALTY) -  Protein phosphatase CheZ from Salmonella typhimurium (strain LT2 / SGSC1412 / ATCC 700720)

Seq: 214 a.a.

Struc: 14 a.a.

Enzyme reactions

• Enzyme class: Chain B: E.C.3.1.3.- - ?????

DOI no: 10.1016/j.jmb.2006.03.050

J Mol Biol 359:624-645 (2006)

PubMed id: 16674976

Crystal structures of beryllium fluoride-free and beryllium fluoride-bound CheY in complex with the conserved C-terminal peptide of CheZ reveal dual binding modes specific to CheY conformation.

J.Guhaniyogi, V.L.Robinson, A.M.Stock.

ABSTRACT

Chemotaxis, the environment-specific swimming behavior of a bacterial cell is controlled by flagellar rotation. The steady-state level of the phosphorylated or activated form of the response regulator CheY dictates the direction of flagellar rotation. CheY phosphorylation is regulated by a fine equilibrium of three phosphotransfer activities: phosphorylation by the kinase CheA, its auto-dephosphorylation and dephosphorylation by its phosphatase CheZ. Efficient dephosphorylation of CheY by CheZ requires two spatially distinct protein-protein contacts: tethering of the two proteins to each other and formation of an active site for dephosphorylation. The former involves interaction of phosphorylated CheY with the small highly conserved C-terminal helix of CheZ (CheZ(C)), an indispensable structural component of the functional CheZ protein. To understand how the CheZ(C) helix, representing less than 10% of the full-length protein, ascertains molecular specificity of binding to CheY, we have determined crystal structures of CheY in complex with a synthetic peptide corresponding to 15 C-terminal residues of CheZ (CheZ(200-214)) at resolutions ranging from 2.0 A to 2.3A. These structures provide a detailed view of the CheZ(C) peptide interaction both in the presence and absence of the phosphoryl analog, BeF3-. Our studies reveal that two different modes of binding the CheZ(200-214) peptide are dictated by the conformational state of CheY in the complex. Our structures suggest that the CheZ(C) helix binds to a "meta-active" conformation of inactive CheY and it does so in an orientation that is distinct from the one in which it binds activated CheY. Our dual binding mode hypothesis provides implications for reverse information flow in CheY and extends previous observations on inherent resilience in CheY-like signaling domains.

Selected figure(s)

Figure 3.
Figure 3. The CheZ[200-214] peptide-CheY interface. Ribbon representation of (a) the F432YZ[200-214] interface and (b) the P2(1)2(1)2YZ[200-214] interface. The side-chains of key contacting residues are illustrated as ball and stick models and hydrophobic contacts are shown as green patches. (c) Relative B-factors of CheZ[200-214] in the CheY-CheZ[200-214] structures. The relative B-factor versus CheZ residue number plot in Click to view the MathML source- [0?wchp=dGLbVlb-zSkzk] -free F432YZ[200-214] is shown in cyan, that in Click to view the MathML source- [0?wchp=dGLbVlb-zSkzk] -bound F432YZ[200-214] in deep blue and that in Click to view the MathML source- [0?wchp=dGLbVlb-zSkzk] -bound P2(1)2(1)2YZ[200-214] in orange. B[residue] is the overall B-factor for each residue and B[CheZ] is the overall B-factor for all atoms of CheZ[200-214] included in the final model. (d) Schematic representation of the CheY-CheZ[200-214] contacts. The F432Z[200-214] primary sequence in cyan and the P2(1)2(1)2Z[200-214] primary sequence in orange are shown on either side of the C-terminal half of CheY, represented in secondary structural elements. Participating residues are highlighted. Hydrophobic contacts are illustrated as continuous grey lines, salt bridges as broken black lines and hydrogen bonds as continuous black lines. The Click to view the MathML source- [0?wchp=dGLbVlb-zSkzk] -free and Click to view the MathML source- [0?wchp=dGLbVlb-zSkzk] -bound F432YZ[200-214] structures solved from crystals grown in Tris (pH 8.4) are used as representatives of F432YZ[200-214] structures in this Figure.

Figure 4.
Figure 4. Ribbon diagrams of CheY-CheZ[C] structures upon superposition of CheY showing different orientations of CheZ[C]. CheY molecules in F432YZ[200-214], P2(1)2(1)2YZ[200-214] and CheY-CheZ[1-214]15 structures are shown in light gray and the respective CheZ[C] helices are shown in cyan, orange and magenta, respectively. The Click to view the MathML source- [0?wchp=dGLbVlb-zSkzk] -free F432YZ[200-214] structure solved from a crystal grown in Tris (pH 8.4) is used in this Figure as a representative of all six F432YZ[200-214] structures.

The above figures are reprinted by permission from Elsevier: J Mol Biol (2006, 359, 624-645) copyright 2006.

Figures were selected by the author.