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Some lessons from the systematic production and structural analysis of soluble (alpha)(beta) T-cell receptors

DOI: 10.1016/j.jim.2009.08.008 DOI Help

Authors: Gijs I. Van Boxel (Cancer Research UK Receptor Structure Research Group, Division of Structural Biology) , Guillaume Stewart-jones (Cancer Research UK Receptor Structure Research Group, Division of Structural Biology) , Samantha Holmes (Cancer Research UK Receptor Structure Research Group, Division of Structural Biology) , Sarah Sainsbury (Oxford Protein Production Facility, Division of Structural Biology) , Dawn Shepherd (Medical Research Council Human Immunology Unit, Weatherall Institute of Molecular Medicine, Nuffield Department of Medicine, University of Oxford) , G. M. A. Gillespie (Medical Research Council Human Immunology Unit, Weatherall Institute of Molecular Medicine, Nuffield Department of Medicine, University of Oxford) , Karl Harlos (Cancer Research UK Receptor Structure Research Group, Division of Structural Biology) , David I. Stuart (Diamond Light Source) , Ray Owens (Oxford Protein Production Facility, Division of Structural Biology) , E. Yvonne Jones (Cancer Research UK Receptor Structure Research Group, Division of Structural Biology)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Journal Of Immunological Methods , VOL 350 , PAGES 14-21

State: Published (Approved)
Published: October 2009

Abstract: T-cell receptors (TCRs) are membrane proteins which recognize antigens with high specificity forming the basis of the cellular immune response. The study of these receptors has been limited by the challenges in expressing sufficient quantities of stable soluble protein. Here we report our systematic approach for generating soluble, ??-TCRs, for X-ray crystallographic studies. By using small-scale expression screens, novel standardized quality control mechanisms and crystallization and imaging robots we were able to add significantly to the current TCR structural database. Our success in crystallizing both isolated TCRs and Major histocompatibility complex (MHC):TCR complexes has provided us with sufficient data to develop focused crystallization screens, which have proved generically useful for the crystallization of this family of proteins and complexes.

Journal Keywords: Cellular Immune Response; Crystallization; High-Throughput; Refolding; Protein Expression

Subject Areas: Biology and Bio-materials


Technical Areas: