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Enhanced Crystallizability by Protein Engineering Approaches: A General Overview

[ Vol. 19 , Issue. 7 ]

Author(s):

Alessia Ruggiero, Giovanni Smaldone, Flavia Squeglia and Rita Berisio   Pages 732 - 742 ( 11 )

Abstract:


The limiting step in macromolecular crystallography is the preparation protein crystals suitable for X-ray diffraction studies. A strong prerequisite for the success of crystallization experiments is the ability to produce monodisperse and properly folded protein samples. Since the production of most protein is usually achieved using recombinant methods, it has become possible to engineer target proteins with increased propensities to form well diffracting crystals. Recent advances in bioinformatics, which takes advantage from an enhanced information in the protein databases, are of enormous help for the design of modified proteins. Based on bioinformatics analyses, the reduction of the structural complexity of proteins or their site-specific mutagenesis has proven to have a dramatic impact on both the yield of heterologous protein expression and its crystallizability.

Therefore, protein engineering represents a valid tool which supports the classical crystallization screenings with a more rational approach. This review describes key methods of protein-engineering and provides a number of examples of their successful use in crystallization.

Scope of Proposed Topic: This Topic is focused on state-of-art protein engineering techniques to increase the propensity of proteins to form crystals with suitable X-ray diffraction properties. Protein engineering methods have proven to be of great help for the crystallization of difficult targets. We herein review molecular biology and chemical methods to help protein crystallization.

Keywords:

Bioinformatics tools, crystallization, fusion protein, protein engineering, protein expression, protein modification, site-directed mutagenesis, diffracting properties, uncrystallized macromolecule, Crystal screens

Affiliation:

Institute of Biostructure and Bioimaging, CNR, Via Mezzocannone 16. I-80134, Napoli, Italy.



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