UV-induced ligand exchange in MHC class I protein crystals

Celie, P.H.N., Toebes, M., Rodenko, B., Ovaa, H., Perrakis, A. and Schumacher, T.N.M. (2009) UV-induced ligand exchange in MHC class I protein crystals. Journal of the American Chemical Society, 131(34), pp. 12298-12304. (doi: 10.1021/ja9037559)

Abstract

High-throughput structure determination of protein−ligand complexes is central in drug development and structural proteomics. To facilitate such high-throughput structure determination we designed an induced replacement strategy. Crystals of a protein complex bound to a photosensitive ligand are exposed to UV light, inducing the departure of the bound ligand, allowing a new ligand to soak in. We exemplify the approach for a class of protein complexes that is especially recalcitrant to high-throughput strategies: the MHC class I proteins. We developed a UV-sensitive, “conditional”, peptide ligand whose UV-induced cleavage in the crystals leads to the exchange of the low-affinity lytic fragments for full-length peptides introduced in the crystallant solution. This “in crystallo” exchange is monitored by the loss of seleno-methionine anomalous diffraction signal of the conditional peptide compared to the signal of labeled MHC β2m subunit. This method has the potential to facilitate high-throughput crystallography in various protein families.

• Item Type: Articles
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Rodenko, Dr Boris
• Authors: Celie, P.H.N., Toebes, M., Rodenko, B., Ovaa, H., Perrakis, A., and Schumacher, T.N.M.
• College/School: College of Medical Veterinary and Life Sciences > School of Infection & Immunity
• Journal Name: Journal of the American Chemical Society
• Publisher: American Chemical Society
• ISSN: 0002-7863
• ISSN (Online): 1520-5126
• Published Online: 05 August 2009
• Copyright Holders: Copyright © 2009 American Chemical Society
• First Published: First published in Journal of the American Chemical Society 131(34):12298-12304
• Publisher Policy: Reproduced in accordance with the copyright policy of the publisher