Membrane Protein Structural Validation by Oriented Sample Solid-State NMR: Diacylglycerol Kinase

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	Membrane Protein Structural Validation by Oriented Sample Solid-State NMR: Diacylglycerol Kinase
	Murray, Dylan T.1,2; Li, Conggang 3; Gao, Philip 4; Qin, Huajun 1,5; Cross, Timothy A.1,2,5
刊名	BIOPHYSICAL JOURNAL
	2014-04-15
卷号	106 期号:8 页码:1559-1569
英文摘要	The validation of protein structures through functional assays has been the norm for many years. Functional assays perform this validation for water-soluble proteins very well, but they need to be performed in the same environment as that used for the structural analysis. This is difficult for membrane proteins that are often structurally characterized in detergent environments, although functional assays for these proteins are most frequently performed in lipid bilayers. Because the structure of membrane proteins is known to be sensitive to the membrane mimetic environment, such functional assays are appropriate for validating the protein construct, but not the membrane protein structure. Here, we compare oriented sample solid-state NMR spectral data of diacylglycerol kinase previously published with predictions of such data from recent structures of this protein. A solution NMR structure of diacylglycerol kinase has been obtained in detergent micelles and three crystal structures have been obtained in a monoolein cubic phase. All of the structures are trimeric with each monomer having three transmembrane and one amphipathic helices. However, the solution NMR structure shows typical perturbations induced by a micelle environment that is reflected in the predicted solid-state NMR resonances from the structural coordinates. The crystal structures show few such perturbations, especially for the wild-type structure and especially for the monomers that do not have significant crystal contacts. For these monomers the predicted and observed data are nearly identical. The thermostabilized constructs do show more perturbations, especially the A41C mutation that introduces a hydrophilic residue into what would be the middle of the lipid bilayer inducing additional hydrogen bonding between trimers. These results demonstrate a general technique for validating membrane protein structures with minimal data obtained from membrane proteins in liquid crystalline lipid bilayers by oriented sample solid-state NMR.
WOS标题词	Science & Technology ; Life Sciences & Biomedicine
类目[WOS]	Biophysics
研究领域[WOS]	Biophysics
关键词[WOS]	M2 PROTON CHANNEL ; ESCHERICHIA-COLI ; LIPIDIC MESOPHASES ; SN-1,2-DIACYLGLYCEROL KINASE ; PHOSPHOLAMBAN PENTAMER ; HELIX ASSOCIATION ; HYBRID SOLUTION ; ENVIRONMENT ; BILAYERS ; TOPOLOGY
收录类别	SCI
语种	英语
WOS记录号	WOS:000334567300007
公开日期	2015-07-14
内容类型	期刊论文
源URL	[http://ir.wipm.ac.cn/handle/112942/1507]
专题	武汉物理与数学研究所_磁共振基础研究部
作者单位	1.Florida State Univ, Natl High Magnet Field Lab, Tallahassee, FL 32306 USA 2.Florida State Univ, Inst Mol Biophys, Tallahassee, FL 32306 USA 3.Chinese Acad Sci, Wuhan Inst Phys & Math, State Key Lab Magnet Resonance & Mol & Atom Phys, Wuhan, Peoples R China 4.Univ Kansas, Del Shankel Struct Biol Ctr, Lawrence, KS 66045 USA 5.Florida State Univ, Dept Chem & Biochem, Tallahassee, FL 32306 USA
推荐引用方式 GB/T 7714	Murray, Dylan T.,Li, Conggang,Gao, Philip,et al. Membrane Protein Structural Validation by Oriented Sample Solid-State NMR: Diacylglycerol Kinase[J]. BIOPHYSICAL JOURNAL,2014,106(8):1559-1569.
APA	Murray, Dylan T.,Li, Conggang,Gao, Philip,Qin, Huajun,&Cross, Timothy A..(2014).Membrane Protein Structural Validation by Oriented Sample Solid-State NMR: Diacylglycerol Kinase.BIOPHYSICAL JOURNAL,106(8),1559-1569.
MLA	Murray, Dylan T.,et al."Membrane Protein Structural Validation by Oriented Sample Solid-State NMR: Diacylglycerol Kinase".BIOPHYSICAL JOURNAL 106.8(2014):1559-1569.