Application of protein engineering to enhance crystallizability and improve crystal properties.
Until recently, protein crystallization has mostly been regarded as a stochastic event over which the investigator has little or no control. With the dramatic technological advances in synchrotron-radiation sources and detectors and the equally impressive progress in crystallographic software, including automated model building and validation, crystallization has increasingly become the rate-limiting step in X-ray diffraction studies of macromolecules. However, with the advent of recombinant methods it has also become possible to engineer target proteins and their complexes for higher propensity to form crystals with desirable X-ray diffraction qualities. As most proteins that are under investigation today are obtained by heterologous overexpression, these techniques hold the promise of becoming routine tools with the potential to transform classical crystallization screening into a more rational high-success-rate approach. This article presents an overview of protein-engineering methods designed to enhance crystallizability and discusses a number of examples of their successful application.
Derewenda
Zygmunt S
ZS
Department of Molecular Physiology and Biological Physics, University of Virginia School of Medicine, Charlottesville, VA 22908-0736, USA. zsd4n@virginia.edu
D000818:Animals; D017360:Arabidopsis / Q000737:chemistry / Q000235:genetics; D029681:Arabidopsis Proteins / Q000737:chemistry / Q000235:genetics; D001426:Bacterial Proteins / Q000737:chemistry; D018360:Crystallography, X-Ray / Q000379:methods*; D004926:Escherichia coli / Q000737:chemistry / Q000235:genetics; D058619:Maltose-Binding Proteins; D008958:Models, Molecular; D033902:Periplasmic Binding Proteins / Q000737:chemistry / Q000235:genetics; D015202:Protein Engineering / Q000379:methods*; D011506:Proteins / Q000737:chemistry*; D000075370:Receptors for Activated C Kinase; D011956:Receptors, Cell Surface / Q000737:chemistry / Q000235:genetics; D012995:Solubility
D029681:Arabidopsis Proteins; D001426:Bacterial Proteins; D058619:Maltose-Binding Proteins; D033902:Periplasmic Binding Proteins; D011506:Proteins; C515163:RACK1 protein, Arabidopsis; D000075370:Receptors for Activated C Kinase; D011956:Receptors, Cell Surface
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