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Helical Membrane Protein Crystallization in the New Era of Electron Cryo-Microscopy

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Structure and Function of Membrane Proteins

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

Abstract

Helical assemblies of proteins, which consist of a two-dimensional lattice of identical subunits arranged with helical symmetry, are a common structural motif in nature. For membrane proteins, crystallization protocols can induce helical arrangements and take advantage of the symmetry found in these assemblies for the structural determination of target proteins. Modern advances in the field of electron cryo-microscopy (cryo-EM), in particular the advent of direct electron detectors, have opened the potential for structure determination of membrane proteins in such assemblies at high resolution. The nature of the symmetry in helical crystals of membrane proteins means that a single image potentially contains enough information for three-dimensional structural determination. With the current direct electron detectors, we have never been closer to making this a reality. Here, we present a protocol detailing the preparation of helical crystals, with an emphasis on further cryo-EM analysis and structural determination of the sarco(endo)plasmic reticulum Ca2+-ATPase in the presence of regulatory subunits such as phospholamban.

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Acknowledgments

Mary D. Hernando and Joseph O. Primeau contributed equally to this work.

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Author notes

  1. Mary D. Hernando and Joseph O. Primeau contributed equally to this work.

Authors and Affiliations

  1. Department of Biochemistry, University of Alberta, Edmonton, AB, Canada

    Mary D. Hernando, Joseph O. Primeau & Howard S. Young

Authors
  1. Mary D. Hernando
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  2. Joseph O. Primeau
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  3. Howard S. Young
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Corresponding author

Correspondence to Howard S. Young .

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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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Hernando, M.D., Primeau, J.O., Young, H.S. (2021). Helical Membrane Protein Crystallization in the New Era of Electron Cryo-Microscopy. 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_10

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  • DOI: https://doi.org/10.1007/978-1-0716-1394-8_10

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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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