A85A_MYCTU
ID A85A_MYCTU Reviewed; 338 AA.
AC P9WQP3; D6MJP3; F2GDG3; P0A4V2; P17944; P17996;
DT 16-APR-2014, integrated into UniProtKB/Swiss-Prot.
DT 16-APR-2014, sequence version 1.
DT 03-AUG-2022, entry version 42.
DE RecName: Full=Diacylglycerol acyltransferase/mycolyltransferase Ag85A;
DE Short=DGAT;
DE EC=2.3.1.122;
DE EC=2.3.1.20;
DE AltName: Full=Acyl-CoA:diacylglycerol acyltransferase;
DE AltName: Full=Antigen 85 complex A;
DE Short=85A;
DE Short=Ag85A;
DE AltName: Full=Fibronectin-binding protein A;
DE Short=Fbps A;
DE Flags: Precursor;
GN Name=fbpA; Synonyms=mpt44; OrderedLocusNames=Rv3804c; ORFNames=MTV026.09c;
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 [GENOMIC DNA].
RC STRAIN=ATCC 35801 / TMC 107 / Erdman;
RX PubMed=2506131; DOI=10.1128/iai.57.10.3123-3130.1989;
RA Borremans M., de Wit L., Volckaert G., Ooms J., de Bruyn J., Huygen K.,
RA van Vooren J.P., Stelandre M., Verhofstadt R., Content J.;
RT "Cloning, sequence determination, and expression of a 32-kilodalton-protein
RT gene of Mycobacterium tuberculosis.";
RL Infect. Immun. 57:3123-3130(1989).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], SUBUNIT, SUBCELLULAR LOCATION, AND
RP DISULFIDE BOND.
RC STRAIN=ATCC 35801 / TMC 107 / Erdman;
RX PubMed=8757831; DOI=10.1128/iai.64.8.3038-3047.1996;
RA Harth G., Lee B.Y., Wang J., Clemens D.L., Horwitz M.A.;
RT "Novel insights into the genetics, biochemistry, and immunocytochemistry of
RT the 30-kilodalton major extracellular protein of Mycobacterium
RT tuberculosis.";
RL Infect. Immun. 64:3038-3047(1996).
RN [3]
RP ERRATUM OF PUBMED:8757831.
RA Harth G., Lee B.Y., Wang J., Clemens D.L., Horwitz M.A.;
RL Infect. Immun. 65:852-852(1997).
RN [4]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=C01;
RX PubMed=20223296; DOI=10.1016/j.meegid.2010.03.003;
RA Chuang P.C., Chen Y.M., Chen H.Y., Jou R.;
RT "Single nucleotide polymorphisms in cell wall biosynthesis-associated genes
RT and phylogeny of Mycobacterium tuberculosis lineages.";
RL Infect. Genet. Evol. 10:459-466(2010).
RN [5]
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., Harris D.E.,
RA Gordon S.V., Eiglmeier K., Gas S., Barry C.E. III, Tekaia F., Badcock K.,
RA Basham D., Brown D., Chillingworth T., Connor R., Davies R.M., Devlin K.,
RA Feltwell T., Gentles S., Hamlin N., Holroyd S., Hornsby T., Jagels K.,
RA Krogh A., McLean J., Moule S., Murphy L.D., Oliver S., Osborne J.,
RA Quail M.A., Rajandream M.A., Rogers J., Rutter S., Seeger K., Skelton S.,
RA Squares S., Squares R., Sulston J.E., Taylor K., Whitehead S.,
RA Barrell B.G.;
RT "Deciphering the biology of Mycobacterium tuberculosis from the complete
RT genome sequence.";
RL Nature 393:537-544(1998).
RN [6]
RP FUNCTION IN THE FIBRONECTIN BINDING.
RX PubMed=3141278; DOI=10.1128/iai.56.12.3046-3051.1988;
RA Abou-Zeid C., Ratliff T.L., Wiker H.G., Harboe M., Bennedsen J., Rook G.A.;
RT "Characterization of fibronectin-binding antigens released by Mycobacterium
RT tuberculosis and Mycobacterium bovis BCG.";
RL Infect. Immun. 56:3046-3051(1988).
RN [7]
RP PROTEIN SEQUENCE OF 44-53.
RX PubMed=2403534; DOI=10.1128/iai.58.1.272-274.1990;
RA Wiker H.G., Sletten K., Nagai S., Harboe M.;
RT "Evidence for three separate genes encoding the proteins of the
RT mycobacterial antigen 85 complex.";
RL Infect. Immun. 58:272-274(1990).
RN [8]
RP FUNCTION AS A MYCOLYLTRANSFERASE, AND DISRUPTION PHENOTYPE.
RX PubMed=12010501; DOI=10.1046/j.1365-2958.2002.02953.x;
RA Puech V., Guilhot C., Perez E., Tropis M., Armitige L.Y., Gicquel B.,
RA Daffe M.;
RT "Evidence for a partial redundancy of the fibronectin-binding proteins for
RT the transfer of mycoloyl residues onto the cell wall arabinogalactan
RT termini of Mycobacterium tuberculosis.";
RL Mol. Microbiol. 44:1109-1122(2002).
RN [9]
RP FUNCTION AS A MYCOLYLTRANSFERASE, AND NOMENCLATURE.
RX PubMed=9162010; DOI=10.1126/science.276.5317.1420;
RA Belisle J.T., Vissa V.D., Sievert T., Takayama K., Brennan P.J.,
RA Besra G.S.;
RT "Role of the major antigen of Mycobacterium tuberculosis in cell wall
RT biogenesis.";
RL Science 276:1420-1422(1997).
RN [10]
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 tuberculosis
RT through an interactome, reactome and genome-scale structural analysis.";
RL BMC Syst. Biol. 2:109-109(2008).
RN [11]
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., Yadav A.K.,
RA Shrivastava P., Marimuthu A., Anand S., Sundaram H., Kingsbury R.,
RA Harsha H.C., Nair B., Prasad T.S., Chauhan D.S., Katoch K., Katoch V.M.,
RA Kumar P., Chaerkady R., Ramachandran S., Dash D., Pandey A.;
RT "Proteogenomic analysis of Mycobacterium tuberculosis by high resolution
RT mass spectrometry.";
RL Mol. Cell. Proteomics 10:M111.011627-M111.011627(2011).
RN [12]
RP FUNCTION AS A DIACYLGLYCEROL ACYLTRANSFERASE, CATALYTIC ACTIVITY,
RP MUTAGENESIS OF SER-169, BIOPHYSICOCHEMICAL PROPERTIES, SUBCELLULAR
RP LOCATION, AND SUBSTRATE SPECIFICITY.
RX PubMed=21819455; DOI=10.1111/j.1365-2958.2011.07792.x;
RA Elamin A.A., Stehr M., Spallek R., Rohde M., Singh M.;
RT "The Mycobacterium tuberculosis Ag85A is a novel diacylglycerol
RT acyltransferase involved in lipid body formation.";
RL Mol. Microbiol. 81:1577-1592(2011).
RN [13]
RP X-RAY CRYSTALLOGRAPHY (2.7 ANGSTROMS) OF 43-338, DISULFIDE BOND, AND
RP SUBUNIT.
RX PubMed=15192106; DOI=10.1074/jbc.m400811200;
RA Ronning D.R., Vissa V., Besra G.S., Belisle J.T., Sacchettini J.C.;
RT "Mycobacterium tuberculosis antigen 85A and 85C structures confirm binding
RT orientation and conserved substrate specificity.";
RL J. Biol. Chem. 279:36771-36777(2004).
CC -!- FUNCTION: The antigen 85 proteins (FbpA, FbpB, FbpC) are responsible
CC for the high affinity of mycobacteria for fibronectin, a large adhesive
CC glycoprotein, which facilitates the attachment of M.tuberculosis to
CC murine alveolar macrophages (AMs). They also help to maintain the
CC integrity of the cell wall by catalyzing the transfer of mycolic acids
CC to cell wall arabinogalactan, and through the synthesis of alpha,alpha-
CC trehalose dimycolate (TDM, cord factor). They catalyze the transfer of
CC a mycoloyl residue from one molecule of alpha,alpha-trehalose
CC monomycolate (TMM) to another TMM, leading to the formation of TDM.
CC FbpA mediates triacylglycerol (TAG) formation with long-chain acyl-CoA
CC as the acyl donor and 1,2-dipalmitoyl-sn-glycerol (1,2-dipalmitin) as
CC the acyl acceptor. It has a preference for C26:0-CoA over C18:1-CoA.
CC {ECO:0000269|PubMed:12010501, ECO:0000269|PubMed:21819455,
CC ECO:0000269|PubMed:3141278, ECO:0000269|PubMed:9162010}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=a 1,2-diacyl-sn-glycerol + an acyl-CoA = a triacyl-sn-glycerol
CC + CoA; Xref=Rhea:RHEA:10868, ChEBI:CHEBI:17815, ChEBI:CHEBI:57287,
CC ChEBI:CHEBI:58342, ChEBI:CHEBI:64615; EC=2.3.1.20;
CC Evidence={ECO:0000269|PubMed:21819455};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=2 alpha,alpha'-trehalose 6-mycolate = alpha,alpha'-trehalose
CC 6,6'-bismycolate + alpha,alpha-trehalose; Xref=Rhea:RHEA:23472,
CC ChEBI:CHEBI:16551, ChEBI:CHEBI:18195, ChEBI:CHEBI:18234;
CC EC=2.3.1.122; Evidence={ECO:0000269|PubMed:21819455};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=306 uM for C26:0-CoA (with 1,2-dipalmitin as the acyl acceptor and
CC at 37 degrees Celsius) {ECO:0000269|PubMed:21819455};
CC KM=390 uM for palmitoleoyl-CoA (with 1,2-dipalmitin as the acyl
CC acceptor and at 37 degrees Celsius) {ECO:0000269|PubMed:21819455};
CC KM=540 uM for C18:0-CoA (with 1,2-dipalmitin as the acyl acceptor and
CC at 37 degrees Celsius) {ECO:0000269|PubMed:21819455};
CC Vmax=3166 nmol/min/mg enzyme (at 37 degrees Celsius)
CC {ECO:0000269|PubMed:21819455};
CC Note=kcat is 55.54 min(-1).;
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:15192106,
CC ECO:0000269|PubMed:8757831}.
CC -!- INTERACTION:
CC PRO_0000000214; PRO_0000009136 [O00602]: FCN1; Xeno; NbExp=2; IntAct=EBI-26878164, EBI-11784425;
CC PRO_0000000214; Q15485: FCN2; Xeno; NbExp=2; IntAct=EBI-26878164, EBI-7468784;
CC PRO_0000000214; O75636: FCN3; Xeno; NbExp=2; IntAct=EBI-26878164, EBI-11786958;
CC PRO_0000000214; PRO_0000017401 [P11226]: MBL2; Xeno; NbExp=2; IntAct=EBI-26878164, EBI-25427940;
CC -!- SUBCELLULAR LOCATION: Secreted, cell wall. Cytoplasm.
CC -!- DISRUPTION PHENOTYPE: Cells lacking this gene produce normally
CC mycoloylated cell wall components. {ECO:0000269|PubMed:12010501}.
CC -!- MISCELLANEOUS: Was identified as a high-confidence drug target.
CC -!- SIMILARITY: Belongs to the mycobacterial A85 antigen family.
CC {ECO:0000305}.
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DR EMBL; M27016; AAA50288.1; -; Genomic_DNA.
DR EMBL; U47335; AAC44295.1; -; Genomic_DNA.
DR EMBL; GQ150314; ADD50052.1; -; Genomic_DNA.
DR EMBL; AL123456; CCP46633.1; -; Genomic_DNA.
DR PIR; H70887; H70887.
DR RefSeq; NP_218321.1; NC_000962.3.
DR RefSeq; WP_003900759.1; NZ_NVQJ01000022.1.
DR PDB; 1SFR; X-ray; 2.70 A; A/B/C=43-338.
DR PDBsum; 1SFR; -.
DR AlphaFoldDB; P9WQP3; -.
DR SMR; P9WQP3; -.
DR IntAct; P9WQP3; 6.
DR STRING; 83332.Rv3804c; -.
DR ESTHER; myctu-a85a; A85-Mycolyl-transferase.
DR MoonProt; P9WQP3; -.
DR PaxDb; P9WQP3; -.
DR DNASU; 886132; -.
DR GeneID; 45427805; -.
DR GeneID; 886132; -.
DR KEGG; mtu:Rv3804c; -.
DR TubercuList; Rv3804c; -.
DR eggNOG; COG0627; Bacteria.
DR OMA; AWARNDP; -.
DR PhylomeDB; P9WQP3; -.
DR BioCyc; MetaCyc:G185E-8100-MON; -.
DR BRENDA; 2.3.1.122; 3445.
DR BRENDA; 2.3.1.20; 3445.
DR Proteomes; UP000001584; Chromosome.
DR GO; GO:0005737; C:cytoplasm; IEA:UniProtKB-SubCell.
DR GO; GO:0005576; C:extracellular region; IDA:CAFA.
DR GO; GO:0009274; C:peptidoglycan-based cell wall; IDA:MTBBASE.
DR GO; GO:0005886; C:plasma membrane; HDA:MTBBASE.
DR GO; GO:0016747; F:acyltransferase activity, transferring groups other than amino-acyl groups; IDA:MTBBASE.
DR GO; GO:0004144; F:diacylglycerol O-acyltransferase activity; IEA:UniProtKB-EC.
DR GO; GO:0050348; F:trehalose O-mycolyltransferase activity; IEA:UniProtKB-EC.
DR GO; GO:0035375; F:zymogen binding; IPI:CAFA.
DR GO; GO:0071769; P:mycolate cell wall layer assembly; IBA:GO_Central.
DR Gene3D; 3.40.50.1820; -; 1.
DR InterPro; IPR029058; AB_hydrolase.
DR InterPro; IPR000801; Esterase-like.
DR InterPro; IPR006311; TAT_signal.
DR Pfam; PF00756; Esterase; 1.
DR SUPFAM; SSF53474; SSF53474; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Acyltransferase; Cell wall; Cytoplasm;
KW Direct protein sequencing; Disulfide bond; Reference proteome; Secreted;
KW Signal; Transferase.
FT SIGNAL 1..43
FT /note="Tat-type signal"
FT /evidence="ECO:0000255"
FT CHAIN 44..338
FT /note="Diacylglycerol acyltransferase/mycolyltransferase
FT Ag85A"
FT /id="PRO_0000000214"
FT REGION 101..111
FT /note="Fibronectin-binding"
FT ACT_SITE 169
FT /note="Nucleophile"
FT /evidence="ECO:0000305"
FT ACT_SITE 273
FT /evidence="ECO:0000250"
FT ACT_SITE 305
FT /evidence="ECO:0000250"
FT BINDING 85..86
FT /ligand="substrate"
FT /evidence="ECO:0000250"
FT BINDING 169
FT /ligand="substrate"
FT /evidence="ECO:0000250"
FT BINDING 197
FT /ligand="substrate"
FT /evidence="ECO:0000250"
FT BINDING 275..278
FT /ligand="substrate"
FT /evidence="ECO:0000250"
FT BINDING 282
FT /ligand="substrate"
FT /evidence="ECO:0000250"
FT BINDING 305..307
FT /ligand="substrate"
FT /evidence="ECO:0000250"
FT DISULFID 130..135
FT /evidence="ECO:0000269|PubMed:15192106,
FT ECO:0000269|PubMed:8757831"
FT MUTAGEN 169
FT /note="S->A: Abolishes diacylglycerol acyltransferase
FT activity."
FT /evidence="ECO:0000269|PubMed:21819455"
FT STRAND 44..46
FT /evidence="ECO:0007829|PDB:1SFR"
FT STRAND 52..58
FT /evidence="ECO:0007829|PDB:1SFR"
FT TURN 59..62
FT /evidence="ECO:0007829|PDB:1SFR"
FT STRAND 63..70
FT /evidence="ECO:0007829|PDB:1SFR"
FT STRAND 78..82
FT /evidence="ECO:0007829|PDB:1SFR"
FT STRAND 89..91
FT /evidence="ECO:0007829|PDB:1SFR"
FT HELIX 93..97
FT /evidence="ECO:0007829|PDB:1SFR"
FT HELIX 100..104
FT /evidence="ECO:0007829|PDB:1SFR"
FT STRAND 110..114
FT /evidence="ECO:0007829|PDB:1SFR"
FT STRAND 129..131
FT /evidence="ECO:0007829|PDB:1SFR"
FT STRAND 134..136
FT /evidence="ECO:0007829|PDB:1SFR"
FT HELIX 140..145
FT /evidence="ECO:0007829|PDB:1SFR"
FT HELIX 147..156
FT /evidence="ECO:0007829|PDB:1SFR"
FT STRAND 160..168
FT /evidence="ECO:0007829|PDB:1SFR"
FT HELIX 170..181
FT /evidence="ECO:0007829|PDB:1SFR"
FT TURN 183..185
FT /evidence="ECO:0007829|PDB:1SFR"
FT STRAND 186..193
FT /evidence="ECO:0007829|PDB:1SFR"
FT HELIX 203..213
FT /evidence="ECO:0007829|PDB:1SFR"
FT HELIX 219..223
FT /evidence="ECO:0007829|PDB:1SFR"
FT HELIX 230..233
FT /evidence="ECO:0007829|PDB:1SFR"
FT TURN 236..239
FT /evidence="ECO:0007829|PDB:1SFR"
FT HELIX 240..246
FT /evidence="ECO:0007829|PDB:1SFR"
FT STRAND 249..253
FT /evidence="ECO:0007829|PDB:1SFR"
FT HELIX 267..289
FT /evidence="ECO:0007829|PDB:1SFR"
FT STRAND 294..298
FT /evidence="ECO:0007829|PDB:1SFR"
FT HELIX 307..316
FT /evidence="ECO:0007829|PDB:1SFR"
FT HELIX 318..325
FT /evidence="ECO:0007829|PDB:1SFR"
SQ SEQUENCE 338 AA; 35686 MW; 57B1CF95D07D52C0 CRC64;
MQLVDRVRGA VTGMSRRLVV GAVGAALVSG LVGAVGGTAT AGAFSRPGLP VEYLQVPSPS
MGRDIKVQFQ SGGANSPALY LLDGLRAQDD FSGWDINTPA FEWYDQSGLS VVMPVGGQSS
FYSDWYQPAC GKAGCQTYKW ETFLTSELPG WLQANRHVKP TGSAVVGLSM AASSALTLAI
YHPQQFVYAG AMSGLLDPSQ AMGPTLIGLA MGDAGGYKAS DMWGPKEDPA WQRNDPLLNV
GKLIANNTRV WVYCGNGKPS DLGGNNLPAK FLEGFVRTSN IKFQDAYNAG GGHNGVFDFP
DSGTHSWEYW GAQLNAMKPD LQRALGATPN TGPAPQGA
