Remember Me
Or use your Academic/Social account:


Or use your Academic/Social account:


You have just completed your registration at OpenAire.

Before you can login to the site, you will need to activate your account. An e-mail will be sent to you with the proper instructions.


Please note that this site is currently undergoing Beta testing.
Any new content you create is not guaranteed to be present to the final version of the site upon release.

Thank you for your patience,
OpenAire Dev Team.

Close This Message


Verify Password:
Verify E-mail:
*All Fields Are Required.
Please Verify You Are Human:
fbtwitterlinkedinvimeoflicker grey 14rssslideshare1
Publisher: American Chemical Society
Languages: English
Types: Article
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.
  • The results below are discovered through our pilot algorithms. Let us know how we are doing!

    • Blundell, T. L.; Jhoti, H.; Abell, C. Nat Rev Drug Discov 2002, 1, 45-54.
    • (2) Bjorkman, P. J.; Saper, M. A.; Samraoui, B.; Bennett, W. S.; Strominger, J. L.; Wiley, D. C. Nature 1987, 329, 512-8.
    • Garcia, K. C.; Adams, E. J. Cell 2005, 122, 333-6.
    • Yewdell, J. W.; Bennink, J. R. Annu Rev Immunol 1999, 17, 51-88.
    • (5) Dudley, M. E.; Nishimura, M. I.; Holt, A. K.; Rosenberg, S. A. J Immunother (1997) 1999, 22, 288-98.
    • (6) Speiser, D. E.; Lienard, D.; Rufer, N.; Rubio-Godoy, V.; Rimoldi, D.; Lejeune, F.; Krieg, A. M.; Cerottini, J. C.; Romero, P. J Clin Invest 2005, 115, 739-46.
    • (7) Borbulevych, O. Y.; Insaidoo, F. K.; Baxter, T. K.; Powell, D. J., Jr.; Johnson, L. A.; Restifo, N. P.; Baker, B. M. J Mol Biol 2007, 372, 1123-36.
    • (8) Hosken, N.; McGowan, P.; Meier, A.; Koelle, D. M.; Sleath, P.; Wagener, F.; Elliott, M.; Grabstein, K.; Posavad, C.; Corey, L. J Virol 2006, 80, 5509-15.
    • (9) Lewinsohn, D. A.; Winata, E.; Swarbrick, G. M.; Tanner, K. E.; Cook, M. S.; Null, M. D.; Cansler, M. E.; Sette, A.; Sidney, J.; Lewinsohn, D. M. PLoS Pathog 2007, 3, 1240-9.
    • (10) Oseroff, C.; Kos, F.; Bui, H. H.; Peters, B.; Pasquetto, V.; Glenn, J.; Palmore, T.; Sidney, J.; Tscharke, D. C.; Bennink, J. R.; Southwood, S.; Grey, H. M.; Yewdell, J. W.; Sette, A. Proc Natl Acad Sci U S A 2005, 102, 13980-5.
    • (11) Sylwester, A. W.; Mitchell, B. L.; Edgar, J. B.; Taormina, C.; Pelte, C.; Ruchti, F.; Sleath, P. R.; Grabstein, K. H.; Hosken, N. A.; Kern, F.; Nelson, J. A.; Picker, L. J. J Exp Med 2005, 202, 673-85.
    • Bouvier, M.; Wiley, D. C. Nat Struct Biol 1998, 5, 377-84.
    • (13) Ljunggren, H. G.; Stam, N. J.; Ohlen, C.; Neefjes, J. J.; Hoglund, P.; Heemels, M. T.; Bastin, J.; Schumacher, T. N.; Townsend, A.; Karre, K.; et al. Nature 1990, 346, 476-80.
    • (14) Schumacher, T. N.; Heemels, M. T.; Neefjes, J. J.; Kast, W. M.; Melief, C. J.; Ploegh, H. L. Cell 1990, 62, 563-7.
    • (18) Rodenko, B.; Toebes, M.; Hadrup, S. R.; van Esch, W. J.; Molenaar, A. M.; Schumacher, T. N.; Ovaa, H. Nat Protoc 2006, 1, 1120-32.
    • (19) Toebes, M.; Coccoris, M.; Bins, A.; Rodenko, B.; Gomez, R.; Nieuwkoop, N. J.; van de Kasteele, W.; Rimmelzwaan, G. F.; Haanen, J. B.; Ovaa, H.; Schumacher, T. N. Nat Med 2006, 12, 246- 51.
    • (20) Bakker, A. H.; Hoppes, R.; Linnemann, C.; Toebes, M.; Rodenko, B.; Berkers, C. R.; Hadrup, S. R.; van Esch, W. J.; Heemskerk, M. H.; Ovaa, H.; Schumacher, T. N. Proc Natl Acad Sci U S A 2008, 105, 3825-30.
    • (21) Newman, J.; Egan, D.; Walter, T. S.; Meged, R.; Berry, I.; Ben Jelloul, M.; Sussman, J. L.; Stuart, D. I.; Perrakis, A. Acta Crystallogr D Biol Crystallogr 2005, 61, 1426-31.
    • (25) Murshudov, G. N.; Vagin, A. A.; Dodson, E. J. Acta Crystallogr D Biol Crystallogr 1997, 53, 240-55.
    • (31) Kontopidis, G.; Andrews, M. J.; McInnes, C.; Cowan, A.; Powers, H.; Innes, L.; Plater, A.; Griffiths, G.; Paterson, D.; Zheleva, D. I.; Lane, D. P.; Green, S.; Walkinshaw, M. D.; Fischer, P. M. Structure 2003, 11, 1537-46.
    • (32) Grotenbreg, G. M.; Roan, N. R.; Guillen, E.; Meijers, R.; Wang, J. H.; Bell, G. W.; Starnbach, M. N.; Ploegh, H. L. Proc Natl Acad Sci U S A 2008, 105, 3831-6.
    • (33) Frickel, E. M.; Sahoo, N.; Hopp, J.; Gubbels, M. J.; Craver, M. P.; Knoll, L. J.; Ploegh, H. L.; Grotenbreg, G. M. J Infect Dis 2008, 198, 1625-33.
    • (34) Grotenbreg, G. M.; Nicholson, M. J.; Fowler, K. D.; Wilbuer, K.; Octavio, L.; Yang, M.; Chakraborty, A. K.; Ploegh, H. L.; Wucherpfennig, K. W. J Biol Chem 2007, 282, 21425-36.
  • No related research data.
  • No similar publications.
  • BioEntity Site Name
    1hhjProtein Data Bank

Share - Bookmark

Funded by projects

Cite this article