title: Identification of an alpha(1-->6) mannopyranosyltransferase (MptA), involved in Corynebacterium glutamicum lipomanann biosynthesis, and identification of its orthologue in Mycobacterium tuberculosis.
abstract: Corynebacterium glutamicum and Mycobacterium tuberculosis share a similar cell wall architecture, and the availability of their genome sequences has enabled the utilization of C. glutamicum as a model for the identification and study of, otherwise essential, mycobacterial genes involved in lipomannan (LM) and lipoarabinomannan (LAM) biosynthesis. We selected the putative glycosyltransferase-Rv2174 from M. tuberculosis and deleted its orthologue NCgl2093 from C. glutamicum. This resulted in the formation of a novel truncated lipomannan (Cg-t-LM) and a complete ablation of LM/LAM biosynthesis. Purification and characterization of Cg-t-LM revealed an overall decrease in molecular mass, a reduction of alpha(1-->6) and alpha(1-->2) glycosidic linkages illustrating a reduced degree of branching compared with wild-type LM. The deletion mutant's biochemical phenotype was fully complemented by either NCgl2093 or Rv2174. Furthermore, the use of a synthetic neoglycolipid acceptor in an in vitro cell-free assay utilizing the sugar donor beta-D-mannopyranosyl-1-monophosphoryl-decaprenol together with the neoglycolipid acceptor alpha-D-Manp-(1-->6)-alpha-D-Manp-O-C8 as a substrate, confirmed NCgl2093 and Rv2174 as an alpha(1-->6) mannopyranosyltransferase (MptA), involved in the latter stages of the biosynthesis of the alpha(1-->6) mannan core of LM. Altogether, these studies have identified a new mannosyltransferase, MptA, and they shed further light on the biosynthesis of LM/LAM in Corynebacterianeae.
authors: School of Biosciences, University of Birmingham, Edgbaston, Birmingham B15 2TT, UK.
authors: Arun K
authors: AK
authors: Mishra
authors: Luke J
authors: LJ
authors: Alderwick
authors: Doris
authors: D
authors: Rittmann
authors: Raju V V
authors: RV
authors: Tatituri
authors: Jerome
authors: J
authors: Nigou
authors: Martine
authors: M
authors: Gilleron
authors: Lothar
authors: L
authors: Eggeling
authors: Gurdyal S
authors: GS
authors: Besra
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism*; D002462:Cell Membrane / Q000201:enzymology; D048230:Corynebacterium glutamicum / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D016680:Genome, Bacterial; D006017:Glycolipids / Q000096:biosynthesis / Q000737:chemistry; D008070:Lipopolysaccharides / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification; D008364:Mannosyltransferases / Q000737:chemistry / Q000378:metabolism*; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D009169:Mycobacterium tuberculosis / Q000201:enzymology*; D017386:Sequence Homology, Amino Acid; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013379:Substrate Specificity
chemical_list: D001426:Bacterial Proteins; D006017:Glycolipids; D008070:Lipopolysaccharides; C064351:lipomannan; D008364:Mannosyltransferases
references: Glycobiology. 2007 Feb;17(2):210-9
references: 17088267
references: Proc Natl Acad Sci U S A. 2006 Sep 12;103(37):13664-9
references: 16945913
references: J Biol Chem. 2007 May 18;282(20):14729-40
references: 17387176
references: Glycobiology. 2007 Jun;17(6):35-56R
references: 17261566
references: J Biol Chem. 1999 Oct 29;274(44):31625-31
references: 10531370
references: J Biol Chem. 2000 Jan 7;275(1):677-84
references: 10617666
references: Biol Chem. 2000 Sep-Oct;381(9-10):899-910
references: 11076021
references: Bioorg Med Chem. 2001 Apr;9(4):815-24
references: 11354664
references: J Biol Chem. 2001 Dec 28;276(52):48854-62
references: 11677227
references: Biochem J. 2002 May 1;363(Pt 3):437-47
references: 11964144
references: Chem Biol. 2002 May;9(5):545-53
references: 12031661
references: Biochem J. 2002 Jul 15;365(Pt 2):441-50
references: 11931640
references: Microbes Infect. 2002 Jul;4(9):945-53
references: 12106787
references: J Biol Chem. 2002 Aug 23;277(34):30635-48
references: 12063260
references: J Biol Chem. 2002 Aug 30;277(35):31335-44
references: 12068013
references: Microbiology. 2002 Oct;148(Pt 10):3029-37
references: 12368436
references: J Biol Chem. 2003 Feb 21;278(8):5513-6
references: 12496255
references: Mol Microbiol. 2003 Apr;48(1):77-84
references: 12657046
references: Tuberculosis (Edinb). 2003;83(1-3):91-7
references: 12758196
references: Biochimie. 2003 Jan-Feb;85(1-2):153-66
references: 12765785
references: Protein Sci. 2003 Jul;12(7):1418-31
references: 12824488
references: J Biol Chem. 2003 Sep 19;278(38):36285-95
references: 12851411
references: Mol Microbiol. 2003 Oct;50(1):69-76
references: 14507364
references: J Biol Chem. 2003 Oct 17;278(42):40842-50
references: 12904287
references: Nucleic Acids Res. 2004 Jan 1;32(Database issue):D138-41
references: 14681378
references: Mol Microbiol. 2004 Feb;51(4):1003-14
references: 14763976
references: Biochem J. 2004 Mar 1;378(Pt 2):589-97
references: 14627436
references: FEMS Microbiol Rev. 2004 May;28(2):225-50
references: 15109786
references: J Biol Chem. 2004 Oct 15;279(42):43540-6
references: 15294902
references: J Biol Chem. 2004 Oct 22;279(43):44847-57
references: 15308633
references: J Biol Chem. 1966 Feb 25;241(4):895-902
references: 4285847
references: J Biol Chem. 1967 Jul 10;242(13):3046-56
references: 4291069
references: J Biol Chem. 1968 Jun 10;243(11):2975-84
references: 4297467
references: J Biol Chem. 1986 Sep 15;261(26):12345-51
references: 3091602
references: J Biol Chem. 1990 Apr 25;265(12):6734-43
references: 2108960
references: Clin Microbiol Rev. 1990 Jul;3(3):227-46
references: 2116939
references: J Biol Chem. 1990 Oct 25;265(30):18200-6
references: 2211696
references: J Biol Chem. 1991 Jul 15;266(20):13217-23
references: 1906464
references: J Bacteriol. 1992 Aug;174(16):5462-5
references: 1644774
references: Science. 1992 Aug 21;257(5073):1055-64
references: 1509256
references: Anal Biochem. 1993 Jan;208(1):57-64
references: 8434796
references: Infect Immun. 1993 Oct;61(10):4173-81
references: 8406806
references: Glycobiology. 1993 Oct;3(5):497-506
references: 8286863
references: J Immunol. 1994 Apr 15;152(8):4070-9
references: 8144972
references: Gene. 1994 Jul 22;145(1):69-73
references: 8045426
references: Biochemistry. 1995 Apr 4;34(13):4257-66
references: 7703239
references: J Biol Chem. 1995 May 26;270(21):12380-9
references: 7759478
references: Annu Rev Biochem. 1995;64:29-63
references: 7574484
references: J Biol Chem. 1997 Jan 3;272(1):117-24
references: 8995236
references: Science. 1997 May 30;276(5317):1420-2
references: 9162010
references: J Biol Chem. 1997 Jul 18;272(29):18460-6
references: 9218490
references: J Biol Chem. 1997 Sep 12;272(37):23094-103
references: 9287310
references: Biochem Soc Trans. 1997 Aug;25(3):845-50
references: 9388559
references: J Biol Chem. 1998 Jan 2;273(1):645-52
references: 9417127
references: Glycobiology. 1998 Feb;8(2):113-20
references: 9451020
references: J Mol Biol. 1999 Feb 5;285(5):2147-60
references: 9925791
references: J Biol Chem. 2005 Feb 18;280(7):5651-63
references: 15546869
references: J Biol Chem. 2005 Sep 16;280(37):32362-71
references: 16040600
references: J Bacteriol. 2005 Nov;187(22):7596-606
references: 16267284
references: J Biol Chem. 2006 Jun 9;281(23):15653-61
references: 16595677
references: J Biol Chem. 2006 Jul 21;281(29):20027-35
references: 16704981
references: J Biol Chem. 2006 Sep 1;281(35):25143-55
references: 16803893
references: J Biol Chem. 2007 Feb 16;282(7):4561-72
references: 17179146