ArticleBiophysical journal2010
LCP-Tm: an assay to measure and understand stability of membrane proteins in a membrane environment.
Article in Biophysical journal, 2010. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 28 papers, 1 of them a synthesis that pooled it.
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Who cites it
28 citing papers in PubMed, 1 synthesis or guideline pooled it.
- A 10-year meta-analysis of membrane protein structural biology: Detergents, membrane mimetics, and structure determination techniques.Biochimica et biophysica acta. Biomembranes · 2021Pooled it
- Effects of Membrane Cholesterol on the Structure and Function of Selected Class A GPCRs─Challenges and Future Perspectives.Biochemistry · 2025Review
- PI(4,5)PAdvances in experimental medicine and biology · 2023Article
- Crystallization of Microbial Rhodopsins.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Evidence that specific interactions play a role in the cholesterol sensitivity of G protein-coupled receptors.Biochimica et biophysica acta. Biomembranes · 2021Article
- Selection of Biophysical Methods for Characterisation of Membrane Proteins.International journal of molecular sciences · 2019Review
- Serial Femtosecond Crystallography of G Protein-Coupled Receptors.Annual review of biophysics · 2018Review
- From Gene to Function: Cell-Free Electrophysiological and Optical Analysis of Ion Pumps in Nanodiscs.Biophysical journal · 2017Article
- An effective thiol-reactive probe for differential scanning fluorimetry with a standard real-time polymerase chain reaction device.Analytical biochemistry · 2016Article
- Nanodisc-Tm: Rapid functional assessment of nanodisc reconstituted membrane proteins by CPM assay.MethodsX · 2016Article
- Methodological advances: the unsung heroes of the GPCR structural revolution.Nature reviews. Molecular cell biology · 2015Review
- Large-scale production and protein engineering of G protein-coupled receptors for structural studies.Frontiers in pharmacology · 2015Review
- Two classes of cholesterol binding sites for the β2AR revealed by thermostability and NMR.Biophysical journal · 2014Article
- Femtosecond crystallography of membrane proteins in the lipidic cubic phase.Philosophical transactions of the Royal Society of London. Series B, Biological sciences · 2014Review
- Quantification of free cysteines in membrane and soluble proteins using a fluorescent dye and thermal unfolding.Nature protocols · 2013Article
- Article
- The GPCR Network: a large-scale collaboration to determine human GPCR structure and function.Nature reviews. Drug discovery · 2013Review
- Fabrication of X-ray compatible microfluidic platforms for protein crystallization.Sensors and actuators. B, Chemical · 2012Article
- Ice breaking in GPCR structural biology.Acta pharmacologica Sinica · 2012Review
- Controlled in meso phase crystallization--a method for the structural investigation of membrane proteins.PloS one · 2012Article
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Authors and funding
4 authors.
Funding
Abstract
Structural and functional studies of membrane proteins are limited by their poor stability outside the native membrane environment. The development of novel methods to efficiently stabilize membrane proteins immediately after purification is important for biophysical studies, and is likely to be critical for studying the more challenging human targets. Lipidic cubic phase (LCP) provides a suitable stabilizing matrix for studying membrane proteins by spectroscopic and other biophysical techniques, including obtaining highly ordered membrane protein crystals for structural studies. We have developed a robust and accurate assay, LCP-Tm, for measuring the thermal stability of membrane proteins embedded in an LCP matrix. In its two implementations, protein denaturation is followed either by a change in the intrinsic protein fluorescence on ligand release, or by an increase in the fluorescence of a thiol-binding reporter dye that measures exposure of cysteines buried in the native structure. Application of the LCP-Tm assay to an engineered human beta2-adrenergic receptor and bacteriorhodopsin revealed a number of factors that increased protein stability in LCP. This assay has the potential to guide protein engineering efforts and identify stabilizing conditions that may improve the chances of obtaining high-resolution structures of intrinsically unstable membrane proteins.
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