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Strategies for crystallizing membrane proteins

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Abstract

Crystallizing membrane proteins remains a challenging endeavor despite the increasing number of membrane protein structures solved by X-ray crystallography. The critical factors in determining the success of the crystallization experiments are the purification and preparation of membrane protein samples. Moreover, there is the added complication that the crystallization conditions must be optimized for use in the presence of detergents although the methods used to crystallize most membrane proteins are, in essence, straightforward applications of standard methodologies for soluble protein crystallization. The roles that detergents play in the stability and aggregation of membrane proteins as well as the colloidal properties of the protein-detergent complexes need to be appreciated and controlledbefore and during the crystallization trials. All X-ray quality crystals of membrane proteins were grown from preparations of detergent-solubilized protein, where the heterogeneous natural lipids from the membrane have been replaced by ahomogeneous detergent environment. It is the preparation of such monodisperse, isotropic solutions of membrane proteins that has allowed the successful application of the standard crystallization methods routinely used on soluble proteins. In this review, the issues of protein purification and sample preparation are addressed as well as the new refinements in crystallization methodologies for membrane proteins. How the physical behavior of the detergent, in the form of micelles or protein-detergent aggregates, affects crystallization and the adaptation of published protocols to new membrane protein systems are also addressed. The general conclusion is that many integral membrane proteins could be crystallized if pure and monodisperse preparations in a suitable detergent system can be prepared.

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Author notes

  1. R. Michael Garavito

    Present address: Department of Biochemistry, Michigan State University, 48824-1319, East Lansing, Michigan

Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, The University Chicago, 920 East 58th Street, 60637, Chicago, Illinois

    R. Michael Garavito

  2. Institut de Biologie Physico-Chimique, 13 rue de Pierre et Marie Curie, F-75005, Paris, France

    Daniel Picot

  3. Department of Pharmacology, University of Pennsylvania School of Medicine, 3620 Hamilton Walk, 19104-6084, Philadelphia, Pennsylvania

    Patrick J. Loll

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  1. R. Michael Garavito
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  2. Daniel Picot
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In memory of Glenn D. Garavito.

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Garavito, R.M., Picot, D. & Loll, P.J. Strategies for crystallizing membrane proteins. J Bioenerg Biomembr 28, 13–27 (1996). https://doi.org/10.1007/BF02150674

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