PDBsum entry 2cia

Sh2-domain

PDB id: 2cia

Contents

Protein chain

98 a.a. *

Ligands

ACE-HIS-ILE-PTR-ASP-GLU-VAL-ALA-ALA-ASP

MPD

IPA

Waters ×103

* Residue conservation analysis

PDB id:

2cia

Name:

Sh2-domain

Title:

Human nck2 sh2-domain in complex with a decaphosphopeptide from translocated intimin receptor (tir) of epec

Structure:

Cytoplasmic protein nck2. Chain: a. Fragment: sh2-domain, residues 284-380. Synonym: nck adaptor protein 2, sh2/sh3 adaptor protein nck-beta, nck-2. Engineered: yes. Translocated intimin receptor. Chain: l. Fragment: phosphopeptide ligand of nck-sh2, residues 471-480.

Source:

Homo sapiens. Human. Organism_taxid: 9606. Expressed in: escherichia coli. Expression_system_taxid: 511693. Synthetic: yes. Escherichia coli. Organism_taxid: 562

Biol. unit:

Dimer (from PDB file)

Resolution:

1.45Å R-factor: 0.150 R-free: 0.170

Authors:

S.Frese,W.-D.Schubert,A.C.Findeis,T.Marquardt,Y.S.Roske, T.E.B.Stradal,D.W.Heinz

Key ref:

S.Frese et al. (2006). The phosphotyrosine peptide binding specificity of Nck1 and Nck2 Src homology 2 domains. J Biol Chem, 281, 18236-18245. PubMed id: 16636066 DOI: 10.1074/jbc.M512917200

Date:

17-Mar-06 Release date: 24-Apr-06

Protein chain

O43639  (NCK2_HUMAN) -  Cytoplasmic protein NCK2 from Homo sapiens

Seq: 380 a.a.

Struc: 98 a.a.*

* PDB and UniProt seqs differ at 1 residue position (black cross)

DOI no: 10.1074/jbc.M512917200

J Biol Chem 281:18236-18245 (2006)

PubMed id: 16636066

The phosphotyrosine peptide binding specificity of Nck1 and Nck2 Src homology 2 domains.

S.Frese, W.D.Schubert, A.C.Findeis, T.Marquardt, Y.S.Roske, T.E.Stradal, D.W.Heinz.

ABSTRACT

Nck proteins are essential Src homology (SH) 2 and SH3 domain-bearing adapters that modulate actin cytoskeleton dynamics by linking proline-rich effector molecules to tyrosine kinases or phosphorylated signaling intermediates. Two mammalian pathogens, enteropathogenic Escherichia coli and vaccinia virus, exploit Nck as part of their infection strategy. Conflicting data indicate potential differences in the recognition specificities of the SH2 domains of the isoproteins Nck1 (Nckalpha) and Nck2 (Nckbeta and Grb4). We have characterized the binding specificities of both SH2 domains and find them to be essentially indistinguishable. Crystal structures of both domains in complex with phosphopeptides derived from the enteropathogenic E. coli protein Tir concur in identifying highly conserved, specific recognition of the phosphopeptide. Differential peptide recognition can therefore not account for the preference of either Nck in particular signaling pathways. Binding studies using sequentially mutated, high affinity phosphopeptides establish the sequence variability tolerated in peptide recognition. Based on this binding motif, we identify potential new binding partners of Nck1 and Nck2 and confirm this experimentally for the Arf-GAP GIT1.

Selected figure(s)

Figure 2.
FIGURE 2. Phosphopeptide complexes of Nck1-SH2 and Nck2-SH2. A, Nck1-SH2 binds Tir12 (yellow), a 12-residue peptide from the EPEC virulence factor Tir. Two N-terminal glutamate residues of the peptide, not resolved in the electron density, are excluded. The electrostatic surface reveals a deep, positively charged (blue) phosphotyrosine binding pocket. B, Nck2-SH2 complexed to Tir8 (green). C, schematic representation of the Nck-SH2/Tir interaction. Chemical bonds of Tir12 are shown in black, and those of residues of Nck are shown in orange. Hydrogen bonds are indicated by dashed lines, hydrophobic interactions by green arcs, and a cation- interaction by a blue dashed line.

Figure 3.
FIGURE 3. Nck1-SH2-Tir12 complex (shades of yellow) superimposed on Nck2-SH2/Tir8 (green). A, the interactions of SH2 domain and peptide are conserved superimposing the peptides. B, the 17 nonconserved residues between Nck1- and Nck2-SH2 (green spheres) are distributed over the SH2 surfaces away from the peptide-protein interface.

The above figures are reprinted by permission from the ASBMB: J Biol Chem (2006, 281, 18236-18245) copyright 2006.

Figures were selected by an automated process.