ID GLFT2_MYCTU Reviewed; 637 AA. AC O53585; L0TFB5; DT 13-JUL-2010, integrated into UniProtKB/Swiss-Prot. DT 01-JUN-1998, sequence version 1. DT 12-SEP-2018, entry version 92. DE RecName: Full=Galactofuranosyltransferase GlfT2 {ECO:0000305}; DE Short=GalTr 2 {ECO:0000305}; DE EC=2.4.1.288 {ECO:0000269|PubMed:18055597, ECO:0000269|PubMed:19571009, ECO:0000305|PubMed:16704275}; DE AltName: Full=Arabinogalactan galactosyltransferase 2 {ECO:0000305}; DE AltName: Full=Galactofuranosylgalactofuranosylrhamnosyl-N-acetylglucosaminyl-diphospho-decaprenol beta-1,5/1,6-galactofuranosyltransferase; DE AltName: Full=Polymerizing galactofuranosyltransferase GlfT2 {ECO:0000305}; GN Name=glfT2 {ECO:0000303|PubMed:18423586, ECO:0000312|EMBL:CCP46637.1}; GN Synonyms=glfT {ECO:0000303|PubMed:11304545, GN ECO:0000303|PubMed:16704275}; OrderedLocusNames=Rv3808c; OS Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv). OC Bacteria; Actinobacteria; Corynebacteriales; Mycobacteriaceae; OC Mycobacterium; Mycobacterium tuberculosis complex. OX NCBI_TaxID=83332; 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 PATHWAY. 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, SUBSTRATE SPECIFICITY, AND PATHWAY. 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, AND BIOPHYSICOCHEMICAL RP 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, CATALYTIC ACTIVITY, 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, CATALYTIC ACTIVITY, AND POLYMER LENGTH CONTROL. RC STRAIN=H37Rv; 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 IDENTIFICATION BY MASS SPECTROMETRY [LARGE SCALE ANALYSIS]. RC STRAIN=ATCC 25618 / H37Rv; RX PubMed=21969609; DOI=10.1074/mcp.M111.011627; RA Kelkar D.S., Kumar D., Kumar P., Balakrishnan L., Muthusamy B., RA Yadav A.K., Shrivastava P., Marimuthu A., Anand S., Sundaram H., RA Kingsbury R., Harsha H.C., Nair B., Prasad T.S., Chauhan D.S., RA Katoch K., Katoch V.M., Kumar P., Chaerkady R., Ramachandran S., RA Dash D., Pandey A.; RT "Proteogenomic analysis of Mycobacterium tuberculosis by high RT resolution mass spectrometry."; RL Mol. Cell. Proteomics 10:M111.011627-M111.011627(2011). RN [10] RP X-RAY CRYSTALLOGRAPHY (2.45 ANGSTROMS) IN COMPLEXES WITH UDP AND RP MANGANESE IONS, MUTAGENESIS OF GLU-300; ASP-371; ASP-372; TRP-399 AND RP HIS-413, BIOPHYSICOCHEMICAL PROPERTIES, COFACTOR, ACTIVE SITE, AND RP 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 galactan polymerization of the CC arabinogalactan (AG) region of the mycolylarabinogalactan- CC peptidoglycan (mAGP) complex, an essential component of the CC mycobacteria cell wall. Thus, successively transfers approximately CC 28 galactofuranosyl (Galf) residues from UDP-galactofuranose (UDP- CC Galf) onto the galactofuranosyl-galactofuranosyl-rhamnosyl-GlcNAc- CC diphospho-decaprenol (Galf-Galf-Rha-GlcNAc-PP-C50) acceptor CC produced by GlfT1, with alternating 1->5 and 1->6 links, forming a CC galactan domain with approximately 30 galactofuranosyl residues. CC {ECO:0000269|PubMed:18055597, ECO:0000269|PubMed:19571009, CC ECO:0000305|PubMed:10934214, ECO:0000305|PubMed:11304545, CC ECO:0000305|PubMed:16704275, ECO:0000305|PubMed:18423586}. 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. {ECO:0000269|PubMed:18055597, CC ECO:0000269|PubMed:19571009, ECO:0000305|PubMed:16704275}. CC -!- COFACTOR: CC Name=Mn(2+); Xref=ChEBI:CHEBI:29035; CC Evidence={ECO:0000305|PubMed:22707726}; CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420; CC Evidence={ECO:0000305|PubMed:22707726}; CC -!- BIOPHYSICOCHEMICAL PROPERTIES: CC Kinetic parameters: CC KM=0.38 mM for UDP-Galf {ECO:0000269|PubMed:18423586, CC ECO:0000269|PubMed:22707726}; CC KM=0.60 mM for beta-D-Galf-(1->5)-beta-D-Galf-(1->6)-beta-D- CC Galf-octyl {ECO:0000269|PubMed:18423586, CC ECO:0000269|PubMed:22707726}; CC KM=1.7 mM for beta-D-Galf-(1->5)-beta-D-Galf-octyl CC {ECO:0000269|PubMed:16704275}; CC KM=0.635 mM for beta-D-Galf-(1->6)-beta-D-Galf-octyl CC {ECO:0000269|PubMed:16704275}; CC KM=0.208 mM for beta-D-Galf-(1->5)-beta-D-Galf-(1->6)-beta-D- CC Galf-octyl {ECO:0000269|PubMed:16704275}; CC KM=0.204 mM for beta-D-Galf-(1->6)-beta-D-Galf-(1->5)-beta-D- CC Galf-octyl {ECO:0000269|PubMed:16704275}; CC Vmax=4.4 umol/min/mg enzyme for galtactose transfer on beta-D- CC Galf-(1->5)-beta-D-Galf-(1->6)-beta-D-Galf-octyl CC {ECO:0000269|PubMed:16704275, ECO:0000269|PubMed:18423586}; CC Note=kcat is 430 min(-1) for galtactose transfer on beta-D-Galf- CC (1->5)-beta-D-Galf-(1->6)-beta-D-Galf-octyl. CC {ECO:0000269|PubMed:18423586, ECO:0000269|PubMed:22707726}; CC -!- PATHWAY: Cell wall biogenesis; cell wall polysaccharide CC biosynthesis. {ECO:0000305|PubMed:11304545, CC ECO:0000305|PubMed:16704275}. CC -!- SUBUNIT: Homotetramer. {ECO:0000269|PubMed:22707726}. CC -!- SUBCELLULAR LOCATION: Cell membrane {ECO:0000269|PubMed:11304545}. CC -!- MISCELLANEOUS: Was identified as a high-confidence drug target. CC {ECO:0000305|PubMed:19099550}. CC -!- SIMILARITY: Belongs to the glycosyltransferase 2 family. CC {ECO:0000305}. 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; AL123456; CCP46637.1; -; Genomic_DNA. DR PIR; D70888; D70888. DR RefSeq; NP_218325.1; NC_000962.3. DR RefSeq; WP_003900761.1; NZ_KK339374.1. 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; -. DR STRING; 83332.Rv3808c; -. DR BindingDB; O53585; -. DR ChEMBL; CHEMBL1075096; -. DR CAZy; GT2; Glycosyltransferase Family 2. DR PaxDb; O53585; -. DR EnsemblBacteria; CCP46637; CCP46637; Rv3808c. DR GeneID; 886136; -. DR KEGG; mtu:Rv3808c; -. DR TubercuList; Rv3808c; -. DR eggNOG; ENOG410602S; Bacteria. DR eggNOG; COG1216; LUCA. DR HOGENOM; HOG000247585; -. DR KO; K16650; -. DR OMA; GWWMCLI; -. DR PhylomeDB; O53585; -. DR BioCyc; MetaCyc:G185E-8104-MONOMER; -. DR BRENDA; 2.4.1.288; 3445. DR UniPathway; UPA00963; -. DR PRO; PR:O53585; -. DR Proteomes; UP000001584; Chromosome. 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:0016740; F:transferase activity; IDA:UniProtKB. DR GO; GO:0016757; F:transferase activity, transferring glycosyl groups; IDA:UniProtKB. DR GO; GO:0035250; F:UDP-galactosyltransferase activity; IDA:MTBBASE. DR GO; GO:0045227; P:capsule polysaccharide biosynthetic process; IEA:UniProtKB-UniPathway. DR GO; GO:0044038; P:cell wall macromolecule biosynthetic process; IDA:UniProtKB. 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: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. DR InterPro; IPR029044; Nucleotide-diphossugar_trans. DR SUPFAM; SSF53448; SSF53448; 2. PE 1: Evidence at protein level; KW 3D-structure; Cell membrane; Cell wall biogenesis/degradation; KW Complete proteome; Glycosyltransferase; Magnesium; Manganese; KW Membrane; Metal-binding; Reference proteome; Transferase. FT CHAIN 1 637 Galactofuranosyltransferase GlfT2. FT /FTId=PRO_0000395356. FT ACT_SITE 372 372 Proton acceptor. FT {ECO:0000305|PubMed:22707726}. FT METAL 256 256 Manganese. {ECO:0000269|PubMed:22707726}. FT METAL 258 258 Manganese. {ECO:0000269|PubMed:22707726}. FT METAL 396 396 Manganese. {ECO:0000269|PubMed:22707726}. FT BINDING 171 171 UDP-Galf. {ECO:0000305|PubMed:22707726}. FT BINDING 200 200 UDP-Galf. {ECO:0000305|PubMed:22707726}. FT BINDING 229 229 UDP-Galf. {ECO:0000305|PubMed:22707726}. FT BINDING 256 256 UDP-Galf. {ECO:0000305|PubMed:22707726}. FT MUTAGEN 300 300 E->S: 1400-fold decrease in transferase FT activity. No effect on affinity for the FT substrate UDP-Galf. FT {ECO:0000269|PubMed:22707726}. FT MUTAGEN 371 371 D->S: Loss of transferase activity. FT {ECO:0000269|PubMed:22707726}. FT MUTAGEN 372 372 D->S: Loss of transferase activity. FT {ECO:0000269|PubMed:22707726}. FT MUTAGEN 399 399 W->S: 1200-fold decrease in transferase FT activity. No effect on affinity for the FT substrate UDP-Galf. FT {ECO:0000269|PubMed:22707726}. FT MUTAGEN 413 413 H->S: 1700-fold decrease in transferase FT activity. No effect on affinity for the FT substrate UDP-Galf. FT {ECO:0000269|PubMed:22707726}. FT HELIX 3 7 {ECO:0000244|PDB:4FIX}. FT STRAND 8 11 {ECO:0000244|PDB:4FIX}. FT HELIX 21 24 {ECO:0000244|PDB:4FIX}. FT TURN 25 27 {ECO:0000244|PDB:4FIX}. FT STRAND 34 36 {ECO:0000244|PDB:4FIX}. FT STRAND 39 42 {ECO:0000244|PDB:4FIX}. FT STRAND 45 48 {ECO:0000244|PDB:4FIX}. FT STRAND 53 55 {ECO:0000244|PDB:4FIX}. FT TURN 59 61 {ECO:0000244|PDB:4FIX}. FT HELIX 65 71 {ECO:0000244|PDB:4FIX}. FT STRAND 75 94 {ECO:0000244|PDB:4FIX}. FT STRAND 100 110 {ECO:0000244|PDB:4FIX}. FT STRAND 112 114 {ECO:0000244|PDB:4FIX}. FT STRAND 116 124 {ECO:0000244|PDB:4FIX}. FT STRAND 128 130 {ECO:0000244|PDB:4FIX}. FT STRAND 133 141 {ECO:0000244|PDB:4FIX}. FT STRAND 143 151 {ECO:0000244|PDB:4FIX}. FT STRAND 162 165 {ECO:0000244|PDB:4FIX}. FT STRAND 168 170 {ECO:0000244|PDB:4FIX}. FT HELIX 172 182 {ECO:0000244|PDB:4FIX}. FT HELIX 186 189 {ECO:0000244|PDB:4FIX}. FT STRAND 192 199 {ECO:0000244|PDB:4FIX}. FT STRAND 201 203 {ECO:0000244|PDB:4FIX}. FT HELIX 205 207 {ECO:0000244|PDB:4FIX}. FT HELIX 211 218 {ECO:0000244|PDB:4FIX}. FT HELIX 219 221 {ECO:0000244|PDB:4FIX}. FT STRAND 222 226 {ECO:0000244|PDB:4FIX}. FT HELIX 231 246 {ECO:0000244|PDB:4FIX}. FT STRAND 250 255 {ECO:0000244|PDB:4FIX}. FT STRAND 257 261 {ECO:0000244|PDB:4FIX}. FT HELIX 264 275 {ECO:0000244|PDB:4FIX}. FT STRAND 276 278 {ECO:0000244|PDB:4FIX}. FT STRAND 281 288 {ECO:0000244|PDB:4FIX}. FT STRAND 300 303 {ECO:0000244|PDB:4FIX}. FT TURN 304 307 {ECO:0000244|PDB:4FIX}. FT STRAND 308 311 {ECO:0000244|PDB:4FIX}. FT TURN 321 323 {ECO:0000244|PDB:4FIX}. FT HELIX 331 334 {ECO:0000244|PDB:4FIX}. FT HELIX 335 337 {ECO:0000244|PDB:4FIX}. FT STRAND 349 353 {ECO:0000244|PDB:4FIX}. FT HELIX 354 360 {ECO:0000244|PDB:4FIX}. FT STRAND 367 370 {ECO:0000244|PDB:4FIX}. FT HELIX 371 381 {ECO:0000244|PDB:4FIX}. FT STRAND 386 396 {ECO:0000244|PDB:4FIX}. FT STRAND 399 402 {ECO:0000244|PDB:4FIX}. FT STRAND 405 407 {ECO:0000244|PDB:4FIY}. FT HELIX 409 424 {ECO:0000244|PDB:4FIX}. FT HELIX 429 431 {ECO:0000244|PDB:4FIX}. FT HELIX 432 448 {ECO:0000244|PDB:4FIX}. FT HELIX 452 466 {ECO:0000244|PDB:4FIX}. FT HELIX 469 474 {ECO:0000244|PDB:4FIX}. FT TURN 475 478 {ECO:0000244|PDB:4FIX}. FT HELIX 479 487 {ECO:0000244|PDB:4FIX}. FT HELIX 491 493 {ECO:0000244|PDB:4FIX}. FT STRAND 494 497 {ECO:0000244|PDB:4FIY}. FT HELIX 499 501 {ECO:0000244|PDB:4FIX}. FT HELIX 518 532 {ECO:0000244|PDB:4FIX}. FT HELIX 538 541 {ECO:0000244|PDB:4FIX}. FT STRAND 545 548 {ECO:0000244|PDB:4FIX}. FT HELIX 550 552 {ECO:0000244|PDB:4FIX}. FT HELIX 555 558 {ECO:0000244|PDB:4FIX}. FT STRAND 562 567 {ECO:0000244|PDB:4FIX}. FT STRAND 574 578 {ECO:0000244|PDB:4FIX}. FT HELIX 581 611 {ECO:0000244|PDB:4FIX}. FT HELIX 613 617 {ECO:0000244|PDB:4FIX}. FT HELIX 619 626 {ECO:0000244|PDB:4FIX}. 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 //