Structural and mechanistic analysis of the membrane-embedded glycosyltransferase WaaA required for lipopolysaccharide synthesis

doi:10.1073/pnas.1119894109

	Structural and mechanistic analysis of the membrane-embedded glycosyltransferase WaaA required for lipopolysaccharide synthesis
	Schmidt, Helgo 1,2; Hansen, Guido 1; Singh, Sonia 2; Hanuszkiewicz, Anna 4; Lindner, Buko 3; Fukase, Koichi 5; Woodard, Ronald W.4; Holst, Otto 2; Hilgenfeld, Rolf 1,6,7; Mamat, Uwe 2
刊名	PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
	2012-04-17
卷号	109 期号:16 页码:6253-6258
关键词	endotoxin GT-B GT-30 monotopic membrane protein
ISSN号	0027-8424
DOI	10.1073/pnas.1119894109
文献子类	Article
英文摘要	WaaA is a key enzyme in the biosynthesis of LPS, a critical component of the outer envelope of Gram-negative bacteria. Embedded in the cytoplasmic face of the inner membrane, WaaA catalyzes the transfer of 3-deoxy-D-manno-oct-2-ulosonic acid (Kdo) to the lipid A precursor of LPS. Here we present crystal structures of the free and CMP-bound forms of WaaA from Aquifex aeolicus, an ancient Gram-negative hyperthermophile. These structures reveal details of the CMP-binding site and implicate a unique sequence motif (GGS/TX(5)GXNXLE) in Kdo binding. In addition, a cluster of highly conserved amino acid residues was identified which represents the potential membrane-attachment and acceptor-substrate binding site of WaaA. A series of site-directed mutagenesis experiments revealed critical roles for glycine 30 and glutamate 31 in Kdo transfer. Our results provide the structural basis of a critical reaction in LPS biosynthesis and allowed the development of a detailed model of the catalytic mechanism of WaaA.
资助项目	European Commission[LSH-2005-037793] ; Deutsche Forschungsgemeinschaft (DFG)[MA 1408/2] ; Deutsche Forschungsgemeinschaft (DFG)[ME 2741/1] ; National Institutes of Health[AI61531] ; DFG[00000000] ; Fonds der Chemischen Industrie[00000000] ; Chinese Academy of Sciences[2010T1S6]
WOS关键词	ESCHERICHIA-COLI ; CRYSTAL-STRUCTURE ; PASTEURELLA-MULTOCIDA ; CATALYTIC MECHANISM ; SIALYLTRANSFERASE ; ACCEPTOR ; COMPLEX ; CMP ; BIOSYNTHESIS ; TRANSFERASE
WOS研究方向	Science & Technology - Other Topics
语种	英语
出版者	NATL ACAD SCIENCES
WOS记录号	WOS:000303246100070
内容类型	期刊论文
源URL	[http://119.78.100.183/handle/2S10ELR8/278116]
专题	中国科学院上海药物研究所
通讯作者	Mamat, Uwe
作者单位	1.Univ Lubeck, Ctr Struct & Cell Biol Med, Inst Biochem, D-23538 Lubeck, Germany; 2.Leibniz Ctr Med & Biosci, Res Ctr Borstel, Div Struct Biochem, D-23845 Borstel, Germany; 3.Leibniz Ctr Med & Biosci, Res Ctr Borstel, Div Immunochem, D-23845 Borstel, Germany; 4.Univ Michigan, Dept Med Chem, Ann Arbor, MI 48109 USA; 5.Osaka Univ, Grad Sch Sci, Dept Chem, Osaka 5600043, Japan; 6.DESY, Lab Struct Biol Infect & Inflammat, D-22603 Hamburg, Germany; 7.Chinese Acad Sci, Shanghai Inst Mat Med, Shanghai 201203, Peoples R China
推荐引用方式 GB/T 7714	Schmidt, Helgo,Hansen, Guido,Singh, Sonia,et al. Structural and mechanistic analysis of the membrane-embedded glycosyltransferase WaaA required for lipopolysaccharide synthesis[J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA,2012,109(16):6253-6258.
APA	Schmidt, Helgo.,Hansen, Guido.,Singh, Sonia.,Hanuszkiewicz, Anna.,Lindner, Buko.,...&Mesters, Jeroen R..(2012).Structural and mechanistic analysis of the membrane-embedded glycosyltransferase WaaA required for lipopolysaccharide synthesis.PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA,109(16),6253-6258.
MLA	Schmidt, Helgo,et al."Structural and mechanistic analysis of the membrane-embedded glycosyltransferase WaaA required for lipopolysaccharide synthesis".PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 109.16(2012):6253-6258.