@article{5d9d013fa73f4f07b61c2b9c5cf4a167,
title = "Ternary structure reveals mechanism of a membrane diacylglycerol kinase",
abstract = "Diacylglycerol kinase catalyses the ATP-dependent conversion of diacylglycerol to phosphatidic acid in the plasma membrane of Escherichia coli. The small size of this integral membrane trimer, which has 121 residues per subunit, means that available protein must be used economically to craft three catalytic and substrate-binding sites centred about the membrane/cytosol interface. How nature has accomplished this extraordinary feat is revealed here in a crystal structure of the kinase captured as a ternary complex with bound lipid substrate and an ATP analogue. Residues, identified as essential for activity by mutagenesis, decorate the active site and are rationalized by the ternary structure. The 3-phosphate of the ATP analogue is positioned for direct transfer to the primary hydroxyl of the lipid whose acyl chain is in the membrane. A catalytic mechanism for this unique enzyme is proposed. The active site architecture shows clear evidence of having arisen by convergent evolution.",
author = "Dianfan Li and Stansfeld, {Phillip J.} and Sansom, {Mark S P} and Aaron Keogh and Lutz Vogeley and Nicole Howe and Lyons, {Joseph A.} and David Aragao and Petra Fromme and Raimund Fromme and Shibom Basu and Ingo Grotjohann and Christopher Kupitz and Kimberley Rendek and Uwe Weierstall and Nadia Zatsepin and Vadim Cherezov and Wei Liu and Sateesh Bandaru and English, {Niall J.} and Cornelius Gati and Anton Barty and Oleksandr Yefanov and Chapman, {Henry N.} and Kay Diederichs and Marc Messerschmidt and S{\'e}bastien Boutet and Williams, {Garth J.} and Seibert, {M. Marvin} and Martin Caffrey",
note = "Funding Information: The work was supported by Science Foundation Ireland grant 12/IA/1255 (M.C.) and 07/SRC/B1160 (J.M.D. MacElroy), the National Institutes of Health Common Fund in Structural Biology grants P50 GM073210 (M.C.), R01 GM095583 (P.F.) and R01 GM108635 (V.C.); National Institute of General Medical Sciences GM75915 (M.C.) and U54 GM094599 (P.F.); and NSF Science and Technology Center award 1231306 (J.C.H.Spence). P.J.S. is supported by BBSRC grant BB/I019855/1. Work in MSPS{\textquoteright}s lab is supported by the Wellcome Trust. C.G. kindly thanks the PIER Helmholtz Graduate School and the Helmholtz Association for financial support. All MD and most DFT simulations were performed using Irish Centre for High-End Computing (ICHEC) facilities, and we thank M. Peters for assistance. We thank C. R. Sanders for providing E. coli strain WH1061. The assistance and support of beamline scientists at the Advanced Photon Source (23-ID), Diamond Light Source (I24), Swiss Light Source (PXII) and LCLS, a National User Facility operated by Stanford University on behalf of the U.S. Department of Energy, Office of Basic Energy Sciences. We thank V. Pye for help with diffraction data collection and preliminary analysis, S. Knapp for advice on kinases, J. Holton for advice on data analysis, and S. Shah for lipid synthesis. J. Spence, E. Xu and R. Stevens are thanked for their encouragement and support.",
year = "2015",
month = dec,
day = "17",
doi = "10.1038/ncomms10140",
language = "English (US)",
volume = "6",
journal = "Nature communications",
issn = "2041-1723",
publisher = "Nature Publishing Group",
}