Biophysical characterization of recombinant proteins: a key to higher structural genomics success.
Hundreds of genomes have been successfully sequenced to date, and the data are publicly available. At the same time, the advances in large-scale expression and purification of recombinant proteins have paved the way for structural genomics efforts. Frequently, however, little is known about newly expressed proteins calling for large-scale protein characterization to better understand their biochemical roles and to enable structure-function relationship studies. In the Structural Genomics Consortium (SGC), we have established a platform to characterize large numbers of purified proteins. This includes screening for ligands, enzyme assays, peptide arrays and peptide displacement in a 384-well format. In this review, we describe this platform in more detail and report on how our approach significantly increases the success rate for structure determination. Coupled with high-resolution X-ray crystallography and structure-guided methods, this platform can also be used toward the development of chemical probes through screening families of proteins against a variety of chemical series and focused chemical libraries.
Vedadi
Masoud
M
Structural Genomics Consortium, University of Toronto, Room 839, MaRS Center, South Tower, Toronto, Ontario, Canada. mvedadi@uhnres.utoronto.ca
Arrowsmith
Cheryl H
CH
Allali-Hassani
Abdellah
A
Senisterra
Guillermo
G
Wasney
Gregory A
GA
D055592:Biophysical Phenomena; D018360:Crystallography, X-Ray; D023281:Genomics / Q000379:methods*; D006801:Humans; D008024:Ligands; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods; D040901:Proteomics / Q000379:methods*; D011994:Recombinant Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D013329:Structure-Activity Relationship
D008024:Ligands; D011994:Recombinant Proteins
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