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

Sangwoo Cho; C Alejandro Velikovsky; Chittoor P Swaminathan; Jon C D Houtman; Lawrence E Samelson; Roy A Mariuzza

doi:10.1038/sj.emboj.7600168

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

Journal article

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Peer reviewed

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

Sangwoo Cho, C Alejandro Velikovsky, Chittoor P Swaminathan, Jon C D Houtman, Lawrence E Samelson and Roy A Mariuzza

The EMBO journal, Vol.23(7), pp.1441-1451

04/07/2004

DOI: 10.1038/sj.emboj.7600168

PMCID: PMC391073

PMID: 15029250

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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.

Adaptor Proteins, Signal Transducing - chemistry

Phosphorylation

Humans

Multiprotein Complexes

Models, Molecular

Molecular Sequence Data

Crystallography, X-Ray

Phosphoproteins - metabolism

Phosphoproteins - chemistry

src Homology Domains

Thermodynamics

Tyrosine - metabolism

Animals

GRB2 Adaptor Protein

Membrane Proteins - chemistry

Adaptor Proteins, Signal Transducing - genetics

Phosphopeptides - genetics

Phosphopeptides - metabolism

Protein Binding

Protein Conformation

Membrane Proteins - metabolism

Mice

Adaptor Proteins, Signal Transducing - metabolism

Binding Sites

Details

Title: Subtitle: Structural basis for differential recognition of tyrosine-phosphorylated sites in the linker for activation of T cells (LAT) by the adaptor Gads
Creators: Sangwoo Cho - Center for Advanced Research in Biotechnology, WM Keck Laboratory for Structural Biology, University of Maryland Biotechnology Institute, Rockville, MD, USA
C Alejandro Velikovsky
Chittoor P Swaminathan
Jon C D Houtman
Lawrence E Samelson
Roy A Mariuzza
Resource Type: Journal article
Publication Details: The EMBO journal, Vol.23(7), pp.1441-1451
DOI: 10.1038/sj.emboj.7600168
PMID: 15029250
PMCID: PMC391073
ISSN: 0261-4189
eISSN: 1460-2075
Grant note: R01 AI036900 / NIAID NIH HHS R37 AI036900 / NIAID NIH HHS AI36900 / NIAID NIH HHS
Language: English
Date published: 04/07/2004
Academic Unit: Microbiology and Immunology; Internal Medicine
Record Identifier: 9984094391002771

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