FADA5_MYCTU
ID FADA5_MYCTU Reviewed; 391 AA.
AC I6XHI4;
DT 30-NOV-2016, integrated into UniProtKB/Swiss-Prot.
DT 03-OCT-2012, sequence version 1.
DT 03-AUG-2022, entry version 75.
DE RecName: Full=Steroid 3-ketoacyl-CoA thiolase {ECO:0000303|PubMed:25482540};
DE EC=2.3.1.16 {ECO:0000269|PubMed:19822655, ECO:0000269|PubMed:25482540};
DE AltName: Full=Acetyl-CoA acetyltransferase FadA5 {ECO:0000303|PubMed:19822655};
DE AltName: Full=Beta-ketoacyl-CoA thiolase {ECO:0000303|PubMed:19822655};
GN Name=fadA5;
GN OrderedLocusNames=Rv3546, LH57_19345 {ECO:0000312|EMBL:AIR16336.1};
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., 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 [2]
RP SEQUENCE REVISION.
RC STRAIN=ATCC 25618 / H37Rv;
RX PubMed=12368430; DOI=10.1099/00221287-148-10-2967;
RA Camus J.-C., Pryor M.J., Medigue C., Cole S.T.;
RT "Re-annotation of the genome sequence of Mycobacterium tuberculosis
RT H37Rv.";
RL Microbiology 148:2967-2973(2002).
RN [3]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 27294 / TMC 102 / H37Rv;
RA Hazbon M.H., Riojas M.A., Damon A.M., Alalade R.O., Cantwell B.J.,
RA Monaco A., King S., Sohrabi A.;
RT "Phylogenetic analysis of Mycobacterial species using whole genome
RT sequences.";
RL Submitted (SEP-2014) to the EMBL/GenBank/DDBJ databases.
RN [4]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, INDUCTION,
RP DISRUPTION PHENOTYPE, PATHWAY, AND SUBSTRATE SPECIFICITY.
RC STRAIN=H37Rv;
RX PubMed=19822655; DOI=10.1128/iai.00893-09;
RA Nesbitt N.M., Yang X., Fontan P., Kolesnikova I., Smith I., Sampson N.S.,
RA Dubnau E.;
RT "A thiolase of Mycobacterium tuberculosis is required for virulence and
RT production of androstenedione and androstadienedione from cholesterol.";
RL Infect. Immun. 78:275-282(2010).
RN [5]
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 [6]
RP X-RAY CRYSTALLOGRAPHY (1.70 ANGSTROMS) OF WILD-TYPE AND MUTANT SER-93 IN
RP COMPLEX WITH SUBSTRATE ANALOGS, FUNCTION, CATALYTIC ACTIVITY,
RP BIOPHYSICOCHEMICAL PROPERTIES, ACTIVE SITE, AND SUBUNIT.
RX PubMed=25482540; DOI=10.1016/j.str.2014.10.010;
RA Schaefer C.M., Lu R., Nesbitt N.M., Schiebel J., Sampson N.S., Kisker C.;
RT "FadA5 a thiolase from Mycobacterium tuberculosis: a steroid-binding pocket
RT reveals the potential for drug development against tuberculosis.";
RL Structure 23:21-33(2015).
CC -!- FUNCTION: Involved in the beta-oxidation of the cholesterol side chain
CC (PubMed:19822655). It is important for utilization of cholesterol as a
CC sole carbon source in vitro and for full virulence in the chronic stage
CC of mouse lung infection (PubMed:19822655). Catalyzes the thiolysis of
CC 3,22-dioxochol-4-en-24-oyl-CoA to yield 3-oxo-4-pregnene-20-carboxyl-
CC CoA (3-OPC-CoA) and acetyl-CoA (PubMed:25482540). Also able to use
CC acetoacetyl-CoA (AcAcCoA) as substrate (PubMed:19822655).
CC {ECO:0000269|PubMed:19822655, ECO:0000269|PubMed:25482540}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=acetyl-CoA + an acyl-CoA = a 3-oxoacyl-CoA + CoA;
CC Xref=Rhea:RHEA:21564, ChEBI:CHEBI:57287, ChEBI:CHEBI:57288,
CC ChEBI:CHEBI:58342, ChEBI:CHEBI:90726; EC=2.3.1.16;
CC Evidence={ECO:0000269|PubMed:19822655, ECO:0000269|PubMed:25482540};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=3-oxochol-4-en-22-oyl-CoA + acetyl-CoA = 3,22-dioxochol-4-en-
CC 24-oyl-CoA + CoA; Xref=Rhea:RHEA:46312, ChEBI:CHEBI:57287,
CC ChEBI:CHEBI:57288, ChEBI:CHEBI:83792, ChEBI:CHEBI:86014;
CC Evidence={ECO:0000269|PubMed:19822655, ECO:0000269|PubMed:25482540};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=3.1 uM for CoA {ECO:0000269|PubMed:25482540};
CC KM=15 uM for CoA {ECO:0000269|PubMed:19822655};
CC KM=11.8 uM for 3,22-dioxochol-4-en-24-oyl-CoA
CC {ECO:0000269|PubMed:25482540};
CC KM=464 uM for AcAcCoA {ECO:0000269|PubMed:19822655};
CC Note=kcat is 0.725 sec(-1) for 3,22-dioxochol-4-en-24-oyl-CoA
CC substrate (PubMed:25482540). kcat is 0.076 sec(-1) for AcAcCoA
CC substrate (PubMed:19822655). kcat is 0.018 sec(-1) for CoA substrate
CC (PubMed:19822655). {ECO:0000269|PubMed:19822655,
CC ECO:0000269|PubMed:25482540};
CC -!- PATHWAY: Steroid metabolism; cholesterol degradation.
CC {ECO:0000269|PubMed:19822655}.
CC -!- SUBUNIT: Dimer of dimers. {ECO:0000269|PubMed:25482540}.
CC -!- INTERACTION:
CC I6XHI4; I6XHI4: fadA5; NbExp=3; IntAct=EBI-16132055, EBI-16132055;
CC -!- INDUCTION: Induced by cholesterol and repressed by KstR.
CC {ECO:0000269|PubMed:19822655}.
CC -!- DISRUPTION PHENOTYPE: Cells lacking this gene display an attenuated
CC disease phenotype with reduced colony-forming units in comparison to
CC the wild-type. This mutant is unable to metabolize cholesterol to
CC androst-4-ene-3,17-dione (AD) and androsta-1,4-diene-3,17-dione (ADD).
CC {ECO:0000269|PubMed:19822655}.
CC -!- SIMILARITY: Belongs to the thiolase-like superfamily. Thiolase family.
CC {ECO:0000305}.
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DR EMBL; AL123456; CCP46368.1; -; Genomic_DNA.
DR EMBL; CP009480; AIR16336.1; -; Genomic_DNA.
DR RefSeq; NP_218063.1; NC_000962.3.
DR RefSeq; WP_003419307.1; NZ_NVQJ01000014.1.
DR PDB; 4UBT; X-ray; 1.70 A; A/B/C/D=1-391.
DR PDB; 4UBU; X-ray; 3.00 A; A/B/C/D/E/F/G/H=1-391.
DR PDB; 4UBV; X-ray; 1.95 A; A/B=1-391.
DR PDB; 4UBW; X-ray; 2.70 A; A/B=1-391.
DR PDB; 5ONC; X-ray; 2.19 A; A/B=1-391.
DR PDBsum; 4UBT; -.
DR PDBsum; 4UBU; -.
DR PDBsum; 4UBV; -.
DR PDBsum; 4UBW; -.
DR PDBsum; 5ONC; -.
DR AlphaFoldDB; I6XHI4; -.
DR SMR; I6XHI4; -.
DR STRING; 83332.Rv3546; -.
DR SwissLipids; SLP:000001007; -.
DR PaxDb; I6XHI4; -.
DR PRIDE; I6XHI4; -.
DR DNASU; 887360; -.
DR GeneID; 45427530; -.
DR GeneID; 887360; -.
DR KEGG; mtu:Rv3546; -.
DR PATRIC; fig|83332.111.peg.3951; -.
DR TubercuList; Rv3546; -.
DR eggNOG; COG0183; Bacteria.
DR HOGENOM; CLU_031026_2_3_11; -.
DR OMA; EWNVDLP; -.
DR PhylomeDB; I6XHI4; -.
DR BioCyc; MetaCyc:G185E-7823-MON; -.
DR UniPathway; UPA01058; -.
DR Proteomes; UP000001584; Chromosome.
DR GO; GO:0003988; F:acetyl-CoA C-acyltransferase activity; IBA:GO_Central.
DR GO; GO:0042802; F:identical protein binding; IPI:IntAct.
DR GO; GO:0006707; P:cholesterol catabolic process; IEA:UniProtKB-UniPathway.
DR GO; GO:0006635; P:fatty acid beta-oxidation; IBA:GO_Central.
DR GO; GO:0010124; P:phenylacetate catabolic process; IBA:GO_Central.
DR CDD; cd00751; thiolase; 1.
DR Gene3D; 3.40.47.10; -; 2.
DR InterPro; IPR002155; Thiolase.
DR InterPro; IPR016039; Thiolase-like.
DR InterPro; IPR020617; Thiolase_C.
DR InterPro; IPR020613; Thiolase_CS.
DR InterPro; IPR020616; Thiolase_N.
DR Pfam; PF02803; Thiolase_C; 1.
DR Pfam; PF00108; Thiolase_N; 1.
DR PIRSF; PIRSF000429; Ac-CoA_Ac_transf; 1.
DR SUPFAM; SSF53901; SSF53901; 2.
DR TIGRFAMs; TIGR01930; AcCoA-C-Actrans; 1.
DR PROSITE; PS00737; THIOLASE_2; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Acyltransferase; Cholesterol metabolism; Lipid degradation;
KW Lipid metabolism; Reference proteome; Steroid metabolism;
KW Sterol metabolism; Transferase; Virulence.
FT CHAIN 1..391
FT /note="Steroid 3-ketoacyl-CoA thiolase"
FT /id="PRO_0000438502"
FT ACT_SITE 93
FT /note="Acyl-thioester intermediate"
FT /evidence="ECO:0000305|PubMed:25482540,
FT ECO:0007744|PDB:4UBV"
FT ACT_SITE 347
FT /note="Proton acceptor"
FT /evidence="ECO:0000305|PubMed:25482540,
FT ECO:0007744|PDB:4UBV"
FT ACT_SITE 377
FT /note="Proton acceptor"
FT /evidence="ECO:0000305|PubMed:25482540"
FT BINDING 151
FT /ligand="CoA"
FT /ligand_id="ChEBI:CHEBI:57287"
FT /evidence="ECO:0000269|PubMed:25482540,
FT ECO:0007744|PDB:4UBT"
FT BINDING 221..223
FT /ligand="CoA"
FT /ligand_id="ChEBI:CHEBI:57287"
FT /evidence="ECO:0000269|PubMed:25482540,
FT ECO:0007744|PDB:4UBT, ECO:0007744|PDB:4UBV"
FT BINDING 246
FT /ligand="CoA"
FT /ligand_id="ChEBI:CHEBI:57287"
FT /evidence="ECO:0000269|PubMed:25482540,
FT ECO:0007744|PDB:4UBT, ECO:0007744|PDB:4UBU,
FT ECO:0007744|PDB:4UBV"
FT BINDING 379
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:25482540,
FT ECO:0007744|PDB:4UBT"
FT STRAND 4..11
FT /evidence="ECO:0007829|PDB:4UBT"
FT TURN 20..23
FT /evidence="ECO:0007829|PDB:4UBT"
FT HELIX 26..42
FT /evidence="ECO:0007829|PDB:4UBT"
FT STRAND 43..45
FT /evidence="ECO:0007829|PDB:4UBW"
FT HELIX 49..51
FT /evidence="ECO:0007829|PDB:4UBT"
FT STRAND 54..58
FT /evidence="ECO:0007829|PDB:4UBT"
FT HELIX 64..66
FT /evidence="ECO:0007829|PDB:4UBT"
FT TURN 67..69
FT /evidence="ECO:0007829|PDB:4UBT"
FT HELIX 70..77
FT /evidence="ECO:0007829|PDB:4UBT"
FT STRAND 85..91
FT /evidence="ECO:0007829|PDB:4UBT"
FT HELIX 92..94
FT /evidence="ECO:0007829|PDB:4UBT"
FT HELIX 95..108
FT /evidence="ECO:0007829|PDB:4UBT"
FT STRAND 113..122
FT /evidence="ECO:0007829|PDB:4UBT"
FT TURN 123..125
FT /evidence="ECO:0007829|PDB:4UBT"
FT TURN 128..132
FT /evidence="ECO:0007829|PDB:4UBT"
FT HELIX 136..139
FT /evidence="ECO:0007829|PDB:4UBT"
FT HELIX 151..162
FT /evidence="ECO:0007829|PDB:4UBT"
FT HELIX 166..185
FT /evidence="ECO:0007829|PDB:4UBT"
FT TURN 186..192
FT /evidence="ECO:0007829|PDB:4UBT"
FT STRAND 196..201
FT /evidence="ECO:0007829|PDB:4UBT"
FT STRAND 207..214
FT /evidence="ECO:0007829|PDB:4UBT"
FT HELIX 225..230
FT /evidence="ECO:0007829|PDB:4UBT"
FT HELIX 242..244
FT /evidence="ECO:0007829|PDB:4UBT"
FT STRAND 249..259
FT /evidence="ECO:0007829|PDB:4UBT"
FT HELIX 260..265
FT /evidence="ECO:0007829|PDB:4UBT"
FT STRAND 272..281
FT /evidence="ECO:0007829|PDB:4UBT"
FT TURN 284..286
FT /evidence="ECO:0007829|PDB:4UBT"
FT HELIX 291..302
FT /evidence="ECO:0007829|PDB:4UBT"
FT HELIX 306..308
FT /evidence="ECO:0007829|PDB:4UBT"
FT STRAND 310..314
FT /evidence="ECO:0007829|PDB:4UBT"
FT HELIX 319..329
FT /evidence="ECO:0007829|PDB:4UBT"
FT HELIX 333..335
FT /evidence="ECO:0007829|PDB:4UBT"
FT HELIX 342..345
FT /evidence="ECO:0007829|PDB:4UBT"
FT TURN 349..351
FT /evidence="ECO:0007829|PDB:4UBT"
FT HELIX 352..367
FT /evidence="ECO:0007829|PDB:4UBT"
FT STRAND 371..378
FT /evidence="ECO:0007829|PDB:4UBT"
FT TURN 379..381
FT /evidence="ECO:0007829|PDB:4UBT"
FT STRAND 382..390
FT /evidence="ECO:0007829|PDB:4UBT"
SQ SEQUENCE 391 AA; 41329 MW; D0B4C64729B00D76 CRC64;
MGYPVIVEAT RSPIGKRNGW LSGLHATELL GAVQKAVVDK AGIQSGLHAG DVEQVIGGCV
TQFGEQSNNI SRVAWLTAGL PEHVGATTVD CQCGSGQQAN HLIAGLIAAG AIDVGIACGI
EAMSRVGLGA NAGPDRSLIR AQSWDIDLPN QFEAAERIAK RRGITREDVD VFGLESQRRA
QRAWAEGRFD REISPIQAPV LDEQNQPTGE RRLVFRDQGL RETTMAGLGE LKPVLEGGIH
TAGTSSQISD GAAAVLWMDE AVARAHGLTP RARIVAQALV GAEPYYHLDG PVQSTAKVLE
KAGMKIGDID IVEINEAFAS VVLSWARVHE PDMDRVNVNG GAIALGHPVG CTGSRLITTA
LHELERTDQS LALITMCAGG ALSTGTIIER I
