PDBsum entry 1r1s

Peptide binding protein

PDB id: 1r1s

Contents

Protein chains

96 a.a. *

Ligands

ACE-PRO-ASP-PTR-GLU-ASN-LEU ×4

SO4 ×7

Waters ×441

* Residue conservation analysis

PDB id:

1r1s

Name:

Peptide binding protein

Title:

Structural basis for differential recognition of tyrosine phosphorylated sites in the linker for activation of t cells (lat) by the adaptor protein gads

Structure:

Grb2-related adaptor protein 2. Chain: a, c, e, g. Fragment: gads-sh2 domain. Synonym: gads protein, growth factor receptor binding protein, grblg, grb-2-like protein, grb2l, hematopoietic cell-associated adaptor protein grpl, grb-2-related monocytic adapter protein, monocytic adapter, mona, adapter protein grid. Engineered: yes. Lat py226 peptide.

Source:

Mus musculus. House mouse. Organism_taxid: 10090. Gene: gads. Expressed in: escherichia coli. Expression_system_taxid: 562. Synthetic: yes. Other_details: the peptide was chemically synthesized

Biol. unit:

Dimer (from PQS)

Resolution:

1.90Å R-factor: 0.217 R-free: 0.259

Authors:

S.Cho,R.A.Mariuzza

Key ref:

S.Cho et al. (2004). Structural basis for differential recognition of tyrosine-phosphorylated sites in the linker for activation of T cells (LAT) by the adaptor Gads. EMBO J, 23, 1441-1451. PubMed id: 15029250 DOI: 10.1038/sj.emboj.7600168

Date:

24-Sep-03 Release date: 28-Sep-04

Protein chains

O89100  (GRAP2_MOUSE) -  GRB2-related adaptor protein 2 from Mus musculus

Seq: 322 a.a.

Struc: 96 a.a.

DOI no: 10.1038/sj.emboj.7600168

EMBO J 23:1441-1451 (2004)

PubMed id: 15029250

Structural basis for differential recognition of tyrosine-phosphorylated sites in the linker for activation of T cells (LAT) by the adaptor Gads.

S.Cho, C.A.Velikovsky, C.P.Swaminathan, J.C.Houtman, L.E.Samelson, R.A.Mariuzza.

ABSTRACT

The transmembrane protein, linker for activation of T cells (LAT), is essential for T-cell activation and development. Phosphorylation of LAT at multiple tyrosines creates binding sites for the adaptors Gads and Grb2, leading to nucleation of multiprotein signaling complexes. Human LAT contains five potential binding sites for Gads, of which only those at Tyr171 and Tyr191 appear necessary for T-cell function. We asked whether Gads binds preferentially to these sites, as differential recognition could assist in assembling defined LAT-based complexes. Measured calorimetrically, Gads-SH2 binds LAT tyrosine phosphorylation sites 171 and 191 with higher affinities than the other sites, with the differences ranging from only several fold weaker binding to no detectable interaction. Crystal structures of Gads-SH2 complexed with phosphopeptides representing sites 171, 191 and 226 were determined to 1.8-1.9 A resolutions. The structures reveal the basis for preferential recognition of specific LAT sites by Gads, as well as for the relatively greater promiscuity of the related adaptor Grb2, whose binding also requires asparagine at position +2 C-terminal to the phosphorylated tyrosine.

Selected figure(s)

Figure 6.
Figure 6 Ribbon diagrams of Gads-SH2 and Grb2 structures. (A) Gads-SH2 in complex with pLAT171. (B) Gads-SH2 in complex with pLAT191. (C) Gads-SH2 in complex with pLAT226. (D) Grb2-SH2 in complex with KPFpYVNV (PDB entry code 1tze) (Rahuel et al, 1996). Secondary structure elements are labeled following the nomenclature for Lck-SH2 (Eck et al, 1993), and are colored as follows: -helices, red and yellow; -strands, green; and loop regions, gold. The N- and C-termini are labeled. The bound peptides are silver, with side chain oxygen and nitrogen atoms colored red and blue, respectively, and phosphorus atoms green.

Figure 8.
Figure 8 Stereo images of Gads-SH2 and Grb2-SH2 structures. (A) View of the +1 binding site in the Gads-SH2/pLAT226 complex. Relevant stretches of the Gads-SH2 polypeptide backbone are cream; labels identify side chains and bound waters (red balls) in the vicinity of +1 Glu of pLAT226 (silver). The side chain of the modeled +1 His residue is green. (B) The +1 binding site in the Grb2-SH2/KPFpYVNV complex (PDB entry code 1tze) (Rahuel et al, 1996). The Grb2-SH2 polypeptide backbone is cream. Side chains and waters (red balls) in the vicinity of +1 Glu of the bound peptide (silver) are labeled; the modeled +1 His is green.

The above figures are reprinted from an Open Access publication published by Macmillan Publishers Ltd: EMBO J (2004, 23, 1441-1451) copyright 2004.

Figures were selected by an automated process.