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UV-induced ligand exchange in MHC class I protein crystals.
- Source :
-
Journal of the American Chemical Society [J Am Chem Soc] 2009 Sep 02; Vol. 131 (34), pp. 12298-304. - Publication Year :
- 2009
-
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 beta2m subunit. This method has the potential to facilitate high-throughput crystallography in various protein families.
- Subjects :
- Amino Acid Sequence
Crystallization
Crystallography, X-Ray
Humans
Ligands
Models, Molecular
Protein Binding radiation effects
Protein Conformation radiation effects
Selenium chemistry
HLA-A2 Antigen chemistry
HLA-A2 Antigen metabolism
Oligopeptides chemistry
Oligopeptides metabolism
Ultraviolet Rays
Subjects
Details
- Language :
- English
- ISSN :
- 1520-5126
- Volume :
- 131
- Issue :
- 34
- Database :
- MEDLINE
- Journal :
- Journal of the American Chemical Society
- Publication Type :
- Academic Journal
- Accession number :
- 19655750
- Full Text :
- https://doi.org/10.1021/ja9037559