4bpd

Structure determination of an integral membrane kinase

spinqualitylabelsall models 4bpd, resolution 3.30Å Show:Asymmetric Unit Biological Assembly Drag the structure with the mouse to rotate   Export Animated Image   Print3D Structural highlights 4bpd is a 6 chain structure with sequence from Ecoli. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance. Ligands: 78M, ZN Activity: Diacylglycerol kinase, with EC number 2.7.1.107 Resources: FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT Function [KDGL_ECOLI] Recycling of diacylglycerol produced during the turnover of membrane phospholipid. Publication Abstract from PubMed 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-A 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. Cell-free expression and in meso crystallisation of an integral membrane kinase for structure determination.,Boland C, Li D, Shah ST, Haberstock S, Dotsch V, Bernhard F, Caffrey M Cell Mol Life Sci. 2014 Jul 11. PMID:25012698 From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. See Also Diacylglycerol kinase

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