ID GLFT2_MYCTU Reviewed; 637 AA. AC O53585; DT 13-JUL-2010, integrated into UniProtKB/Swiss-Prot. DT 01-JUN-1998, sequence version 1. DT 03-APR-2013, entry version 61. DE RecName: Full=Galactofuranosyl transferase GlfT2; DE Short=GalTr; DE EC=2.4.1.288; DE AltName: Full=Beta-D-(1->5)galactofuranosyltransferase; DE AltName: Full=Beta-D-(1->6)galactofuranosyltransferase; DE AltName: Full=Galactofuranosylgalactofuranosylrhamnosyl-N-acetylglucosaminyl-diphospho-decaprenol beta-1,5/1,6-galactofuranosyltransferase; DE AltName: Full=UDP-Galf:alpha-3-L-rhamnosyl-alpha-D-GlcNAc-pyrophosphate polyprenol, UDP-galactofuranosyl transferase; GN Name=glfT2; OrderedLocusNames=Rv3808c; OS Mycobacterium tuberculosis. OC Bacteria; Actinobacteria; Actinobacteridae; Actinomycetales; OC Corynebacterineae; Mycobacteriaceae; Mycobacterium; OC Mycobacterium tuberculosis complex. OX NCBI_TaxID=1773; RN [1] RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. RC STRAIN=ATCC 25618 / H37Rv; RX PubMed=9634230; DOI=10.1038/31159; RA Cole S.T., Brosch R., Parkhill J., Garnier T., Churcher C.M., RA Harris D.E., Gordon S.V., Eiglmeier K., Gas S., Barry C.E. III, RA Tekaia F., Badcock K., Basham D., Brown D., Chillingworth T., RA Connor R., Davies R.M., Devlin K., Feltwell T., Gentles S., Hamlin N., RA Holroyd S., Hornsby T., Jagels K., Krogh A., McLean J., Moule S., RA Murphy L.D., Oliver S., Osborne J., Quail M.A., Rajandream M.A., RA Rogers J., Rutter S., Seeger K., Skelton S., Squares S., Squares R., RA Sulston J.E., Taylor K., Whitehead S., Barrell B.G.; RT "Deciphering the biology of Mycobacterium tuberculosis from the RT complete genome sequence."; RL Nature 393:537-544(1998). RN [2] RP FUNCTION AS A GALACTOFURANOSYLTRANSFERASE. RX PubMed=10934214; DOI=10.1074/jbc.M006875200; RA Mikusova K., Yagi T., Stern R., McNeil M.R., Besra G.S., Crick D.C., RA Brennan P.J.; RT "Biosynthesis of the galactan component of the mycobacterial cell RT wall."; RL J. Biol. Chem. 275:33890-33897(2000). RN [3] RP FUNCTION AS A GALACTOFURANOSYLTRANSFERASE, SUBCELLULAR LOCATION, AND RP NOMENCLATURE. RC STRAIN=ATCC 25618 / H37Rv; RX PubMed=11304545; DOI=10.1074/jbc.M102022200; RA Kremer L., Dover L.G., Morehouse C., Hitchin P., Everett M., RA Morris H.R., Dell A., Brennan P.J., McNeil M.R., Flaherty C., RA Duncan K., Besra G.S.; RT "Galactan biosynthesis in Mycobacterium tuberculosis. Identification RT of a bifunctional UDP-galactofuranosyltransferase."; RL J. Biol. Chem. 276:26430-26440(2001). RN [4] RP FUNCTION AS A GALACTOFURANOSYLTRANSFERASE, CATALYTIC ACTIVITY, RP BIOPHYSICOCHEMICAL PROPERTIES, AND SUBSTRATE SPECIFICITY. RX PubMed=16704275; DOI=10.1021/ja058254d; RA Rose N.L., Completo G.C., Lin S.J., McNeil M., Palcic M.M., RA Lowary T.L.; RT "Expression, purification, and characterization of a RT galactofuranosyltransferase involved in Mycobacterium tuberculosis RT arabinogalactan biosynthesis."; RL J. Am. Chem. Soc. 128:6721-6729(2006). RN [5] RP IDENTIFICATION AS A DRUG TARGET [LARGE SCALE ANALYSIS]. RX PubMed=19099550; DOI=10.1186/1752-0509-2-109; RA Raman K., Yeturu K., Chandra N.; RT "targetTB: a target identification pipeline for Mycobacterium RT tuberculosis through an interactome, reactome and genome-scale RT structural analysis."; RL BMC Syst. Biol. 2:109-109(2008). RN [6] RP FUNCTION AS A GALACTOFURANOSYLTRANSFERASE, CATALYTIC ACTIVITY, AND RP BIOPHYSICOCHEMICAL PROPERTIES. RX PubMed=18423586; DOI=10.1016/j.carres.2008.03.023; RA Rose N.L., Zheng R.B., Pearcey J., Zhou R., Completo G.C., RA Lowary T.L.; RT "Development of a coupled spectrophotometric assay for GlfT2, a RT bifunctional mycobacterial galactofuranosyltransferase."; RL Carbohydr. Res. 343:2130-2139(2008). RN [7] RP FUNCTION AS A GALACTOFURANOSYLTRANSFERASE, AND SUBSTRATE SPECIFICITY. RX PubMed=18055597; DOI=10.1128/JB.01326-07; RA Belanova M., Dianiskova P., Brennan P.J., Completo G.C., Rose N.L., RA Lowary T.L., Mikusova K.; RT "Galactosyl transferases in mycobacterial cell wall synthesis."; RL J. Bacteriol. 190:1141-1145(2008). RN [8] RP FUNCTION, AND ELONGATION LENGTH CONTROL. RX PubMed=19571009; DOI=10.1073/pnas.0901407106; RA May J.F., Splain R.A., Brotschi C., Kiessling L.L.; RT "A tethering mechanism for length control in a processive carbohydrate RT polymerization."; RL Proc. Natl. Acad. Sci. U.S.A. 106:11851-11856(2009). RN [9] RP X-RAY CRYSTALLOGRAPHY (2.45 ANGSTROMS) IN COMPLEX WITH SUBSTRATE AND RP MANGANESE IONS, MUTAGENESIS OF GLU-300; ASP-371; ASP-372; TRP-399 AND RP HIS-413, BIOPHYSICOCHEMICAL PROPERTIES, COFACTOR, AND SUBUNIT. RX PubMed=22707726; DOI=10.1074/jbc.M112.347484; RA Wheatley R.W., Zheng R.B., Richards M.R., Lowary T.L., Ng K.K.; RT "Tetrameric structure of the GlfT2 galactofuranosyltransferase reveals RT a scaffold for the assembly of mycobacterial Arabinogalactan."; RL J. Biol. Chem. 287:28132-28143(2012). CC -!- FUNCTION: Involved in the polymerization of the arabinogalactan CC (AG) region of the mycolylarabinogalactan-peptidoglycan (mAGP) CC complex, an essential component the mycobacteria cell wall, CC through successively beta-D-(1->5) and beta-D-(1->6)- CC galactofuranosyltransferases activities. It transfers the CC galactofuranosyl (Galf) unit from UDP-galactofuranosyl (UDP-Galf) CC onto the galactofuranosyl-rhamnosyl-GlcNAc-pyrophosphoryl- CC undecaprenyl (Galf-Rha-GlcNAc-PP-C55), yielding CC polygalactofuranosyl-rhamnosyl-GlcNAc-pyrophosphoryl-undecaprenyl CC ((Galf)x-Rha-GlcNAc-PP-C55). It is able to transfer Galf onto CC beta-D-(1->5) or beta-D-(1->6) linked acceptor, but has a stronger CC affinity for beta-D-(1->6) acceptor. CC -!- CATALYTIC ACTIVITY: 28 UDP-alpha-D-galactofuranose + beta-D- CC galactofuranosyl-(1->5)-beta-D-galactofuranosyl-(1->4)-alpha-L- CC rhamnopyranosyl-(1->3)-N-acetyl-alpha-D-glucosaminyl-diphospho- CC trans,octacis-decaprenol = 28 UDP + (beta-D-galactofuranosyl- CC (1->5)-beta-D-galactofuranosyl-(1->6))(14)-beta-D- CC galactofuranosyl-(1->5)-beta-D-galactofuranosyl-(1->4)-alpha-L- CC rhamnopyranosyl-(1->3)-N-acetyl-alpha-D-glucosaminyl-diphospho- CC trans,octacis-decaprenol. CC -!- COFACTOR: Manganese or magnesium. CC -!- BIOPHYSICOCHEMICAL PROPERTIES: CC Kinetic parameters: CC KM=0.38 mM for UDP-Galf; CC KM=0.38 mM for UDP-Galf (at pH 7.9); CC Vmax=4.4 umol/min/mg enzyme; CC -!- PATHWAY: Cell wall biogenesis; cell wall polysaccharide CC biosynthesis. CC -!- SUBUNIT: Dimer of dimers. CC -!- SUBCELLULAR LOCATION: Membrane. CC -!- MISCELLANEOUS: Was identified as a high-confidence drug target. CC -!- SIMILARITY: Belongs to the glycosyltransferase 2 family. CC --------------------------------------------------------------------------- CC Copyrighted by the UniProt Consortium, see https://www.uniprot.org/terms CC Distributed under the Creative Commons Attribution (CC BY 4.0) License CC --------------------------------------------------------------------------- DR EMBL; BX842584; CAA17872.1; -; Genomic_DNA. DR PIR; D70888; D70888. DR RefSeq; NP_218325.1; NC_000962.3. DR RefSeq; YP_006517305.1; NC_018143.1. DR PDB; 4FIX; X-ray; 2.45 A; A/B=1-637. DR PDB; 4FIY; X-ray; 3.10 A; A/B=1-637. DR PDBsum; 4FIX; -. DR PDBsum; 4FIY; -. DR ProteinModelPortal; O53585; -. DR SMR; O53585; 1-629. DR STRING; 83332.Rv3808c; -. DR CAZy; GT2; Glycosyltransferase Family 2. DR PRIDE; O53585; -. DR GeneID; 886136; -. DR GenomeReviews; AL123456_GR; Rv3808c. DR KEGG; mtu:Rv3808c; -. DR KEGG; mtv:RVBD_3808c; -. DR PATRIC; 18157074; VBIMycTub87468_4239. DR TubercuList; Rv3808c; -. DR eggNOG; COG1216; -. DR HOGENOM; HOG000247585; -. DR KO; K16650; -. DR OMA; LCLEYST; -. DR ProtClustDB; CLSK872249; -. DR BioCyc; MetaCyc:MONOMER-15256; -. DR UniPathway; UPA00963; -. DR BindingDB; O53585; -. DR ChEMBL; CHEMBL1075096; -. DR GO; GO:0005829; C:cytosol; IDA:MTBBASE. DR GO; GO:0005886; C:plasma membrane; IDA:MTBBASE. DR GO; GO:0035496; F:lipopolysaccharide-1,5-galactosyltransferase activity; IDA:MTBBASE. DR GO; GO:0008921; F:lipopolysaccharide-1,6-galactosyltransferase activity; IDA:MTBBASE. DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW. DR GO; GO:0045227; P:capsule polysaccharide biosynthetic process; IEA:UniProtKB-UniPathway. DR GO; GO:0071555; P:cell wall organization; IDA:UniProtKB. DR GO; GO:0070592; P:cell wall polysaccharide biosynthetic process; IDA:MTBBASE. DR GO; GO:0040007; P:growth; IMP:MTBBASE. DR GO; GO:0009103; P:lipopolysaccharide biosynthetic process; IDA:MTBBASE. DR GO; GO:0071769; P:mycolate cell wall layer assembly; IDA:MTBBASE. DR GO; GO:0052573; P:UDP-D-galactose metabolic process; IDA:MTBBASE. PE 1: Evidence at protein level; KW 3D-structure; Cell wall biogenesis/degradation; Complete proteome; KW Glycosyltransferase; Magnesium; Manganese; Membrane; Metal-binding; KW Reference proteome; Transferase. FT CHAIN 1 637 Galactofuranosyl transferase GlfT2. FT /FTId=PRO_0000395356. FT ACT_SITE 272 272 Proton acceptor (Probable). FT METAL 256 256 Manganese. FT METAL 258 258 Manganese. FT METAL 396 396 Manganese. FT BINDING 171 171 Substrate. FT BINDING 200 200 Substrate. FT BINDING 229 229 Substrate. FT BINDING 256 256 Substrate. FT MUTAGEN 300 300 E->S: Almost identical transferase FT activity than wild-type. FT MUTAGEN 371 371 D->S: Loss of transferase activity. FT MUTAGEN 372 372 D->S: Loss of transferase activity. FT MUTAGEN 399 399 W->S: Almost identical transferase FT activity than wild-type. FT MUTAGEN 413 413 H->S: Almost identical transferase FT activity than wild-type. FT HELIX 3 7 FT STRAND 8 11 FT HELIX 21 24 FT TURN 25 27 FT STRAND 34 36 FT STRAND 39 42 FT STRAND 45 48 FT STRAND 53 55 FT TURN 59 61 FT HELIX 65 71 FT STRAND 75 94 FT STRAND 100 110 FT STRAND 112 114 FT STRAND 116 124 FT STRAND 128 130 FT STRAND 133 141 FT STRAND 143 151 FT STRAND 162 165 FT STRAND 168 170 FT HELIX 172 182 FT HELIX 186 189 FT STRAND 192 199 FT STRAND 201 203 FT HELIX 205 207 FT HELIX 211 218 FT HELIX 219 221 FT STRAND 222 226 FT HELIX 231 246 FT STRAND 250 255 FT STRAND 257 261 FT HELIX 264 275 FT STRAND 276 278 FT STRAND 281 288 FT STRAND 300 303 FT TURN 304 307 FT STRAND 308 311 FT TURN 321 323 FT HELIX 331 334 FT HELIX 335 337 FT STRAND 349 353 FT HELIX 354 360 FT STRAND 367 370 FT HELIX 371 381 FT STRAND 386 396 FT STRAND 399 402 FT STRAND 405 407 FT HELIX 409 424 FT HELIX 429 431 FT HELIX 432 448 FT HELIX 452 466 FT HELIX 469 474 FT TURN 475 478 FT HELIX 479 487 FT HELIX 491 493 FT STRAND 494 497 FT HELIX 499 501 FT HELIX 518 532 FT HELIX 538 541 FT STRAND 545 548 FT HELIX 550 552 FT HELIX 555 558 FT STRAND 562 567 FT STRAND 574 578 FT HELIX 581 611 FT HELIX 613 617 FT HELIX 619 626 SQ SEQUENCE 637 AA; 71507 MW; D7C9AB28B5005B66 CRC64; MSELAASLLS RVILPRPGEP LDVRKLYLEE STTNARRAHA PTRTSLQIGA ESEVSFATYF NAFPASYWRR WTTCKSVVLR VQVTGAGRVD VYRTKATGAR IFVEGHDFTG TEDQPAAVET EVVLQPFEDG GWVWFDITTD TAVTLHSGGW YATSPAPGTA NIAVGIPTFN RPADCVNALR ELTADPLVDQ VIGAVIVPDQ GERKVRDHPD FPAAAARLGS RLSIHDQPNL GGSGGYSRVM YEALKNTDCQ QILFMDDDIR LEPDSILRVL AMHRFAKAPM LVGGQMLNLQ EPSHLHIMGE VVDRSIFMWT AAPHAEYDHD FAEYPLNDNN SRSKLLHRRI DVDYNGWWTC MIPRQVAEEL GQPLPLFIKW DDADYGLRAA EHGYPTVTLP GAAIWHMAWS DKDDAIDWQA YFHLRNRLVV AAMHWDGPKA QVIGLVRSHL KATLKHLACL EYSTVAIQNK AIDDFLAGPE HIFSILESAL PQVHRIRKSY PDAVVLPAAS ELPPPLHKNK AMKPPVNPLV IGYRLARGIM HNLTAANPQH HRRPEFNVPT QDARWFLLCT VDGATVTTAD GCGVVYRQRD RAKMFALLWQ SLRRQRQLLK RFEEMRRIYR DALPTLSSKQ KWETALLPAA NQEPEHG //