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Highlights from recently determined structures of membrane proteins: a focus on channels and transporters.

ABSTRACT: After decades of absent or lackluster growth, recent years have at long last witnessed an exponential growth in the number of novel membrane protein structures determined. Every single achievement has had a tremendous impact on the scientific community, providing an unprecedented wealth of information that typically only an atomic resolution structure can contribute to our molecular understanding of how a protein functions. Presented here is a review of some of the most exciting novel structures of channels and transporters determined by X-ray crystallography in the last two years, and a discussion of their analogies, differences and mechanistic implications.

SUBMITTER: Sciara G

PROVIDER: S-EPMC3392460 | biostudies-literature | 2012 Aug

REPOSITORIES: biostudies-literature

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Highlights from recently determined structures of membrane proteins: a focus on channels and transporters.

Sciara Giuliano G Mancia Filippo F

Current opinion in structural biology 20120401 4

After decades of absent or lackluster growth, recent years have at long last witnessed an exponential growth in the number of novel membrane protein structures determined. Every single achievement has had a tremendous impact on the scientific community, providing an unprecedented wealth of information that typically only an atomic resolution structure can contribute to our molecular understanding of how a protein functions. Presented here is a review of some of the most exciting novel structures ...[more]

PMID: 22472602

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Project description:BackgroundMembrane proteins (MPs) play key roles in signal transduction. However, understanding their function at a molecular level is mostly hampered by the lack of protein in suitable amount and quality. Despite impressive developments in the expression of prokaryotic MPs, eukaryotic MP production has lagged behind and there is a need for new expression strategies. In a pilot study, we produced a Drosophila glutamate receptor specifically in the eyes of transgenic flies, exploiting the naturally abundant membrane stacks in the photoreceptor cells (PRCs). Now we address the question whether the PRCs also process different classes of medically relevant target MPs which were so far notoriously difficult to handle with conventional expression strategies.Principal findingsWe describe the homologous and heterologous expression of 10 different targets from the three major MP classes--G protein-coupled receptors (GPCRs), transporters and channels in Drosophila eyes. PRCs offered an extraordinary capacity to produce, fold and accommodate massive amounts of MPs. The expression of some MPs reached similar levels as the endogenous rhodopsin, indicating that the PRC membranes were almost unsaturable. Expression of endogenous rhodopsin was not affected by the target MPs and both could coexist in the membrane stacks. Heterologous expression levels reached about 270 to 500 pmol/mg total MP, resulting in 0.2-0.4 mg purified target MP from 1 g of fly heads. The metabotropic glutamate receptor and human serotonin transporter--both involved in synaptic transmission--showed native pharmacological characteristics and could be purified to homogeneity as a prerequisite for further studies.SignificanceWe demonstrate expression in Drosophila PRCs as an efficient and inexpensive tool for the large scale production of functional eukaryotic MPs. The fly eye system offers a number of advantages over conventional expression systems and paves the way for in-depth analyses of eukaryotic MPs that have so far not been accessible to biochemical and biophysical studies.

Dataset Information

Highlights from recently determined structures of membrane proteins: a focus on channels and transporters.

Publications

Highlights from recently determined structures of membrane proteins: a focus on channels and transporters.

Similar Datasets

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