International
Tables for
Crystallography
Volume F
Crystallography of biological macromolecules
Edited by E. Arnold, D. M. Himmel and M. G. Rossmann

International Tables for Crystallography (2012). Vol. F, ch. 4.4, p. 140

Section 4.4.1. Introduction

K. H. Choia*

aDepartment of Biochemistry and Molecular Biology, 6.614C Basic Science, The University of Texas Medical Branch,University Blvd, Galveston, TX 77555–0647, USA
Correspondence e-mail: kychoi@utmb.edu

4.4.1. Introduction

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Structural-genomics projects have contributed to major developments in automation, miniaturization and process integration in X-ray crystallography. In the structural-genomics approach, multiple open reading frames from a genome are separately cloned and expressed without prior knowledge of the structure or function of the encoded proteins. Essential steps in a typical project include: (1) bioinformatics analysis of genome sequences for potential targets, (2) gene amplification by PCR (polymerase chain reaction) and subcloning into an appropriate expression vector, (3) protein expression and purification, (4) protein crystallization, (5) X-ray data collection, and (6) data processing and structure determination. Structural-genomics projects typically operate under a `lowest hanging fruit' philosophy, pursuing structural targets that prove to be the most amenable to crystallization. However, for individual investigators working on specific problems, it is often necessary to focus on a particular protein. Many of the protocols developed in high-throughput (HT) structural genomics can also be adapted for parallel production of multiple constructs of a single protein target which is difficult to crystallize. This chapter summarizes recent developments in HT X-ray crystallography approaches and their application to parallel production of a single protein (Gräslund, Nordlund et al., 2008[link]; Manjasetty et al., 2008[link]; Sharff & Jhoti, 2003[link]).

References

Gräslund, S., Nordlund, P. et al. (2008). Protein production and purification. Nat. Methods, 5, 135–146.
Manjasetty, B. A., Turnbull, A. P., Panjikar, S., Bussow, K. & Chance, M. R. (2008). Automated technologies and novel techniques to accelerate protein crystallography for structural genomics. Proteomics, 8, 612–625.
Sharff, A. & Jhoti, H. (2003). High-throughput crystallography to enhance drug discovery. Curr. Opin. Chem. Biol. 7, 340–345.








































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