Application of the use of high-throughput technologies to the determination of protein structures of bacterial and viral pathogens

M. J. Fogg (1) , P. Alzari (2) , M. Bahar (3) , I. Bertini (4) , J.-M. Betton (1) , W. P. Burmeister (5) , C. Cambillau (6) , B. Canard (6) , M. Carrondo (7) , M. Coll (8) , S. Daenke (3) , O. Dym (9) , M.-P. Egloff (6) , F. J. Enguita (7) , A. Geerlof (10) , A. Haouz (2) , T. A. Jones (11) , Q. Ma (10) , S. N. Manicka (9) , M. Migliardi (4) , P. Nordlund (12) , R. J. Owens (3) , Y. Peleg (9) , G. Schneider (13) , R. Schnell (13) , D. I. Stuart (3) , N. Tarbouriech (5) , T. Unge (11) , A. J. Wilkinson (1) , M. Wilmanns (10) , K. S. Wilson (1) , O. Zimhony (9) , J. M. Grimes (3)

1 University of York [York, UK]
2 Biochimie Structurale
3 The Wellcome Trust Centre for Human Genetics [Oxford]
4 CIRMMP - Consorzio Interuniversitario Risonanze Magnetiche di Metallo Proteine
5 EMBL - European Molecular Biology Laboratory [Grenoble]
6 AFMB - Architecture et fonction des Macromolécules Biologiques - UMR 6098
7 ITQB - Instituto de Tecnologia Química e Biológica António Xavier
8 Institut de Biologia Molecular de Barcelona
9 Weizmann Institute of Science [Rehovot, Israël]
10 EMBL - European Molecular Biology Laboratory [Hamburg]
11 BMC - Biomedical Center = Biomedicinskt centrum [Uppsala, Sueden]
12 Stockholm University
13 Karolinska Institutet [Stockholm]

The Structural Proteomics In Europe (SPINE) programme is aimed at the development and implementation of high-throughput technologies for the efficient structure determination of proteins of biomedical importance, such as those of bacterial and viral pathogens linked to human health. Despite the challenging nature of some of these targets, 175 novel pathogen protein structures (approximately 220 including complexes) have been determined to date. Here the impact of several technologies on the structural determination of proteins from human pathogens is illustrated with selected examples, including the parallel expression of multiple constructs, the use of standardized refolding protocols and optimized crystallization screens.

Keywords: automation; miniaturization; cloning; expression; viruses; bacteria

Domains: Cristallography; Structural Biology [q-bio.BM]; Biochemistry, Molecular Biology; Bioinformatics [q-bio.QM]; Biological Physics [physics.bio-ph]; Cellular Biology

M. J. Fogg, P. Alzari, M. Bahar, I. Bertini, J.-M. Betton, et al.. Application of the use of high-throughput technologies to the determination of protein structures of bacterial and viral pathogens. Acta crystallographica Section D : Structural biology [1993-..], 2006, 62 (10), pp.1196-1207. ⟨10.1107/S0907444906030915⟩. ⟨pasteur-03139239⟩