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Structure Determination of Membrane Proteins Using X-Ray Crystallography

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Structure and Function of Membrane Proteins

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2302))

Abstract

Membrane proteins serve essential roles in all aspects of life and make up roughly one-third of all genomes from prokaryotes to eukaryotes. Their responsibilities include mediating cell signaling, nutrient import, waste export, cellular communication, trafficking, and immunity. For their critical role in many cellular processes, membrane proteins serve as targets for up to 50% of drugs currently on the market and remain primary targets in new therapeutics being developed. Despite their importance and abundance in nature, only ~1% of structures in the Protein Data Bank are of transmembrane proteins. This discrepancy can be directly attributed to the biochemical properties of membrane proteins and the difficulty in producing sufficient yields for structural studies or the difficulty in growing well-ordered crystals. Here, we present methods from our work that outline our general pipeline from cloning to structure determination of membrane proteins, with a focus on using X-ray crystallography, which still yields ~90% of all structures being deposited into the Protein Data Bank.

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Acknowledgements

We would like to acknowledge funding supporting this work through grants GM127884 (N.N.), GM127896 (N.N.), and GM132024 (E.B., T32 trainee) from the National Institute of General Medical Sciences (NIGMS) and AI148103 (C.O., T32 trainee) from the National Institute of Allergy and Infectious Disease (NIAID).

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Authors and Affiliations

  1. Markey Center for Structural Biology, Department of Biological Sciences, Purdue Institute of Inflammation, Immunology and Infectious Disease, Purdue University, West Lafayette, IN, USA

    Evan Billings, Karl Lundquist, Claire Overly, Karthik Srinivasan & Nicholas Noinaj

  2. Department of Biological Sciences, Purdue University, West Lafayette, IN, USA

    Evan Billings, Karl Lundquist, Claire Overly, Karthik Srinivasan & Nicholas Noinaj

  3. noinajlab, Purdue University, West Lafayette, IN, USA

    Nicholas Noinaj

Authors
  1. Evan Billings
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  2. Karl Lundquist
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  3. Claire Overly
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  4. Karthik Srinivasan
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  5. Nicholas Noinaj
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Corresponding author

Correspondence to Nicholas Noinaj .

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Editors and Affiliations

  1. School of Biological Sciences and School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, GA, USA

    Ingeborg Schmidt-Krey

  2. School of Physics, Georgia Institute of Technology, Atlanta, GA, USA

    James C. Gumbart

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Billings, E., Lundquist, K., Overly, C., Srinivasan, K., Noinaj, N. (2021). Structure Determination of Membrane Proteins Using X-Ray Crystallography. In: Schmidt-Krey, I., Gumbart, J.C. (eds) Structure and Function of Membrane Proteins. Methods in Molecular Biology, vol 2302. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1394-8_7

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  • DOI: https://doi.org/10.1007/978-1-0716-1394-8_7

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1393-1

  • Online ISBN: 978-1-0716-1394-8

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