ArticleCellular and molecular life sciences : CMLS2014
Cell-free expression and in meso crystallisation of an integral membrane kinase for structure determination.
Article in Cellular and molecular life sciences : CMLS, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Who cites it
12 citing papers in PubMed.
- Solid-state NMR of membrane peptides and proteins in the lipid cubic phase.Biophysical journal · 2025Article
- Cell-Free Gene Expression: Methods and Applications.Chemical reviews · 2025Review
- Reconstitution of a Reversible Membrane Switch via Prenylation by One-Pot Cell-Free Expression.ACS synthetic biology · 2023Article
- Applications of Cell-Free Synthesized Membrane Protein Precipitates.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Cell-Free Synthesis Strategies to Probe Co-translational Folding of Proteins Within Lipid Membranes.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Interfaces Between Alpha-helical Integral Membrane Proteins: Characterization, Prediction, and Docking.Computational and structural biotechnology journal · 2019Article
- LILBID and nESI: Different Native Mass Spectrometry Techniques as Tools in Structural Biology.Journal of the American Society for Mass Spectrometry · 2019Article
- From Gene to Function: Cell-Free Electrophysiological and Optical Analysis of Ion Pumps in Nanodiscs.Biophysical journal · 2017Article
- The cubicon method for concentrating membrane proteins in the cubic mesophase.Nature protocols · 2017Article
- Cell-free methods to produce structurally intact mammalian membrane proteins.Scientific reports · 2016Article
- Ternary structure reveals mechanism of a membrane diacylglycerol kinase.Nature communications · 2015Article
- A comprehensive review of the lipid cubic phase or in meso method for crystallizing membrane and soluble proteins and complexes.Acta crystallographica. Section F, Structural biology communications · 2015Review
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7 authors.
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Abstract
Membrane proteins are key elements in cell physiology and drug targeting, but getting a high-resolution structure by crystallographic means is still enormously challenging. Novel strategies are in big demand to facilitate the structure determination process that will ultimately hasten the day when sequence information alone can provide a three-dimensional model. Cell-free or in vitro expression enables rapid access to large quantities of high-quality membrane proteins suitable for an array of applications. Despite its impressive efficiency, to date only two membrane proteins produced by the in vitro approach have yielded crystal structures. Here, we have analysed synergies of cell-free expression and crystallisation in lipid mesophases for generating an X-ray structure of the integral membrane enzyme diacylglycerol kinase to 2.28-Å resolution. The quality of cellular and cell-free-expressed kinase samples has been evaluated systematically by comparing (1) spectroscopic properties, (2) purity and oligomer formation, (3) lipid content and (4) functionality. DgkA is the first membrane enzyme crystallised based on cell-free expression. The study provides a basic standard for the crystallisation of cell-free-expressed membrane proteins and the methods detailed here should prove generally useful and contribute to accelerating the pace at which membrane protein structures are solved.
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