Current trends in α-helical membrane protein crystallization: An update

Department of Biochemistry, University of Oxford, South Parks Road, Oxford OX1 3QU, United Kingdom.
Protein Science (Impact Factor: 2.85). 09/2012; 21(9):1358-65. DOI: 10.1002/pro.2122
Source: PubMed


α-Helical membrane proteins (MPs) are the targets for many pharmaceutical drugs and play important roles in human physiology. In recent years, significant progress has been made in determining their atomic structure using X-ray crystallography. However, a major bottleneck in MP crystallography still remains, namely, the identification of conditions that give crystals that are suitable for structural determination. In 2008, we undertook an analysis of the crystallization conditions for 121 α-helical MPs to design a rationalized sparse matrix crystallization screen, MemGold. We now report an updated analysis that includes a further 133 conditions. The results reveal the current trends in α-helical MP crystallization with notable differences since 2008. The updated information has been used to design new crystallization and additive screens that should prove useful for both initial crystallization scouting and subsequent crystal optimization.

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Available from: Joanne L Parker, Oct 24, 2014
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    • "In contrast, the space between protein units in type II crystals is usually large enough to accommodate the original detergent belt at the expense of stable protein contacts, which likely contributes to the difficulty in obtaining high-quality crystals for diffraction experiments. Among the many detergents available, n-dodecyl-b-D-maltoside (bDM) provides the largest number of successful crystallization trials (Parker and Newstead, 2012). Photosystem II (PSII) is the water-plastoquinone oxidoreductase of oxygenic photosynthesis. "
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    • "The advantages of the statistical design included a comparable success rate to other screens with a smaller number of chemicals, with a more straightforward path towards optimization than a random screen owing to the repetition of specific chemicals within the screen (Tran et al., 2004). More recent examples of this same approach of data mining and creation of screens to encapsulate the results can be found in the Morpheus screen (Gorrec, 2009) and the MemGold screens (Newstead et al., 2008; Parker & Newstead, 2012). 4.4.2. "
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    ABSTRACT: While crystallization historically predates crystallography, it is a critical step for the crystallographic process. The rich history of crystallization and how that history influences current practices is described. The tremendous impact of crystallization screens on the field is discussed.
    Acta Crystallographica Section F Structural Biology and Crystallization Communications 07/2014; 70(Pt 7):835-53. DOI:10.1107/S2053230X1401262X · 0.53 Impact Factor
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    ABSTRACT: As we appreciate the importance of stabilising membrane proteins, the barriers towards their structure determination are being broken down. This change in mindset comes hand-in-hand with more effort placed on developing methods focused at screening for membrane proteins which are naturally stable in detergent solution or improving those that are not so. In practice, however, it is not easy to decide the best strategy to monitor and improve detergent stability, requiring a decision-making process that can be even more difficult for those new to the field. In this review we outline the importance of membrane protein stability with discussions of the stabilisation strategies applied in context with the use of crystallisation scaffolds and the different types of crystallisation methods themselves. Where possible we also highlight areas that we think could push this field forward with emerging technologies, such as X-ray free electron lasers (X-feL), which could have a big impact on the membrane protein structural biology community. We hope this review will serve as a useful guide for those striving to solve structures of both pro- and eukaryotic membrane proteins.
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