James Brown1, Thomas S. Walter1, L. Carter1, Nicola G. A. Abrescia1, A.R. Aricescu1, Thil Batuwangala1, Louise E. Bird1, Nick R. Brown2, Philip P. Chamberlain1, Simon J. Davis3, Е. О. Дубинина2, Jane Endicott2, Janet A. Fennelly3, Robert J. C. Gilbert1, Maria Harkiolaki4, Wai‐Ching Hon1, Fiona C. Kimberley4, Christopher Love1, Erika J. Mancini1, Raquel Manso-Sancho3, C.E. Nichols1, R.A. Robinson1, Geoff Sutton1, N. Schueller2, Mark C. Sleeman3, Guillaume Stewart‐Jones1, Mai Tuyet Vuong3, Julie P. I. Welburn2, Z. Zhang5, D.K. Stammers1, Raymond J. Owens1, E. Yvonne Jones1, Karl Harlos1, David I. Stuart1
1Oxford Protein Production Facility and The Division of Structural Biology, The Henry Wellcome Building of Genomic Medicine, Roosevelt Drive, Headington, Oxford OX3 7BN, UK
2Laboratory of Molecular Biophysics, Department of Biochemistry, University of Oxford, The Rex Richards Building, South Parks Road, Oxford OX1 3QU, UK
3Nuffield Department of Clinical Medicine, University of Oxford, John Radcliffe Hospital, Oxford, OX3 9DU, UK
4The Weatherall Institute of Molecular Medicine, John Radcliffe Hospital, Headington, Oxford OX3 9DS, UK
5The Dyson Perrins Laboratory, Oxford Centre for Molecular Sciences, University of Oxford, South Parks Road, Oxford OX1 3QY, UK
Tóm tắt
An initial tranche of results from day-to-day use of a robotic system for setting up 100 nl-scale vapour-diffusion sitting-drop protein crystallizations has been surveyed. The database of over 50 unrelated samples represents a snapshot of projects currently at the stage of crystallization trials in Oxford research groups and as such encompasses a broad range of proteins. The results indicate that the nanolitre-scale methodology consistently identifies more crystallization conditions than traditional hand-pipetting-style methods; however, in a number of cases successful scale-up is then problematic. Crystals grown in the initial 100 nl-scale drops have in the majority of cases allowed useful characterization of X-ray diffraction, either in-house or at synchrotron beamlines. For a significant number of projects, full X-ray diffraction data sets have been collected to 3 Å resolution or better (either in-house or at the synchrotron) from crystals grown at the 100 nl scale. To date, five structures have been determined by molecular replacement directly from such data and a further three from scale-up of conditions established at the nanolitre scale.
