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Genetic Construct Design and Recombinant Protein Expression for Structural Biology

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Structure-Based Drug Discovery

Part of the book series: Methods in Molecular Biology ((MIMB,volume 841))

Abstract

Obtaining diffraction quality crystals is frequently an iterative process which traditionally has involved screening large numbers of crystallization conditions. Due to advances in high-throughput gene engineering, recombinant expression, and purification, the protein of interest has now become one of the many variables routinely investigated during crystallization trials. As such, construct design is a critical step in the path toward successful crystallization. In this chapter will we address construct design strategies frequently employed to improve the solution and crystallization behavior of proteins. Topics covered include choosing a recombinant expression system and reducing disorder through truncations and surface mutagenesis. Also covered are strategies to reduce heterogeneity from posttranslational modifications, impurities, and aggregation.

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Authors and Affiliations

  1. Centocor R&D Inc., San Diego, CA, USA

    Suzanne C. Edavettal, Michael J. Hunter & Ronald V. Swanson

Authors
  1. Suzanne C. Edavettal
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  2. Michael J. Hunter
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  3. Ronald V. Swanson
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Correspondence to Ronald V. Swanson .

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Editors and Affiliations

  1. Trius Therapeutics, Nancy Ridge Dr. 6310, San Diego, 92121, California, USA

    Leslie W. Tari

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Edavettal, S.C., Hunter, M.J., Swanson, R.V. (2012). Genetic Construct Design and Recombinant Protein Expression for Structural Biology. In: Tari, L. (eds) Structure-Based Drug Discovery. Methods in Molecular Biology, vol 841. Humana Press. https://doi.org/10.1007/978-1-61779-520-6_2

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  • DOI: https://doi.org/10.1007/978-1-61779-520-6_2

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