BACG_BACSU
ID BACG_BACSU Reviewed; 259 AA.
AC P39644;
DT 01-FEB-1995, integrated into UniProtKB/Swiss-Prot.
DT 01-FEB-1995, sequence version 1.
DT 03-AUG-2022, entry version 137.
DE RecName: Full=NADPH-dependent reductase BacG {ECO:0000305};
DE EC=1.3.1.- {ECO:0000269|PubMed:20052993, ECO:0000269|PubMed:22765234, ECO:0000269|PubMed:23519407};
DE AltName: Full=Bacilysin biosynthesis oxidoreductase YwfH;
DE AltName: Full=H2HPP reductase {ECO:0000305};
GN Name=bacG {ECO:0000303|PubMed:22765234};
GN Synonyms=ywfH {ECO:0000303|PubMed:20052993}; OrderedLocusNames=BSU37680;
GN ORFNames=ipa-86r;
OS Bacillus subtilis (strain 168).
OC Bacteria; Firmicutes; Bacilli; Bacillales; Bacillaceae; Bacillus.
OX NCBI_TaxID=224308;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=168;
RX PubMed=7934828; DOI=10.1111/j.1365-2958.1993.tb01963.x;
RA Glaser P., Kunst F., Arnaud M., Coudart M.P., Gonzales W., Hullo M.-F.,
RA Ionescu M., Lubochinsky B., Marcelino L., Moszer I., Presecan E.,
RA Santana M., Schneider E., Schweizer J., Vertes A., Rapoport G., Danchin A.;
RT "Bacillus subtilis genome project: cloning and sequencing of the 97 kb
RT region from 325 degrees to 333 degrees.";
RL Mol. Microbiol. 10:371-384(1993).
RN [2]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=168;
RX PubMed=9384377; DOI=10.1038/36786;
RA Kunst F., Ogasawara N., Moszer I., Albertini A.M., Alloni G., Azevedo V.,
RA Bertero M.G., Bessieres P., Bolotin A., Borchert S., Borriss R.,
RA Boursier L., Brans A., Braun M., Brignell S.C., Bron S., Brouillet S.,
RA Bruschi C.V., Caldwell B., Capuano V., Carter N.M., Choi S.-K.,
RA Codani J.-J., Connerton I.F., Cummings N.J., Daniel R.A., Denizot F.,
RA Devine K.M., Duesterhoeft A., Ehrlich S.D., Emmerson P.T., Entian K.-D.,
RA Errington J., Fabret C., Ferrari E., Foulger D., Fritz C., Fujita M.,
RA Fujita Y., Fuma S., Galizzi A., Galleron N., Ghim S.-Y., Glaser P.,
RA Goffeau A., Golightly E.J., Grandi G., Guiseppi G., Guy B.J., Haga K.,
RA Haiech J., Harwood C.R., Henaut A., Hilbert H., Holsappel S., Hosono S.,
RA Hullo M.-F., Itaya M., Jones L.-M., Joris B., Karamata D., Kasahara Y.,
RA Klaerr-Blanchard M., Klein C., Kobayashi Y., Koetter P., Koningstein G.,
RA Krogh S., Kumano M., Kurita K., Lapidus A., Lardinois S., Lauber J.,
RA Lazarevic V., Lee S.-M., Levine A., Liu H., Masuda S., Mauel C.,
RA Medigue C., Medina N., Mellado R.P., Mizuno M., Moestl D., Nakai S.,
RA Noback M., Noone D., O'Reilly M., Ogawa K., Ogiwara A., Oudega B.,
RA Park S.-H., Parro V., Pohl T.M., Portetelle D., Porwollik S.,
RA Prescott A.M., Presecan E., Pujic P., Purnelle B., Rapoport G., Rey M.,
RA Reynolds S., Rieger M., Rivolta C., Rocha E., Roche B., Rose M., Sadaie Y.,
RA Sato T., Scanlan E., Schleich S., Schroeter R., Scoffone F., Sekiguchi J.,
RA Sekowska A., Seror S.J., Serror P., Shin B.-S., Soldo B., Sorokin A.,
RA Tacconi E., Takagi T., Takahashi H., Takemaru K., Takeuchi M.,
RA Tamakoshi A., Tanaka T., Terpstra P., Tognoni A., Tosato V., Uchiyama S.,
RA Vandenbol M., Vannier F., Vassarotti A., Viari A., Wambutt R., Wedler E.,
RA Wedler H., Weitzenegger T., Winters P., Wipat A., Yamamoto H., Yamane K.,
RA Yasumoto K., Yata K., Yoshida K., Yoshikawa H.-F., Zumstein E.,
RA Yoshikawa H., Danchin A.;
RT "The complete genome sequence of the Gram-positive bacterium Bacillus
RT subtilis.";
RL Nature 390:249-256(1997).
RN [3]
RP INDUCTION, AND PATHWAY.
RC STRAIN=168 / 61884;
RX PubMed=12372825; DOI=10.1074/jbc.m208722200;
RA Inaoka T., Takahashi K., Ohnishi-Kameyama M., Yoshida M., Ochi K.;
RT "Guanine nucleotides guanosine 5'-diphosphate 3'-diphosphate and GTP co-
RT operatively regulate the production of an antibiotic bacilysin in Bacillus
RT subtilis.";
RL J. Biol. Chem. 278:2169-2176(2003).
RN [4]
RP FUNCTION, CATALYTIC ACTIVITY, AND PATHWAY.
RX PubMed=20052993; DOI=10.1021/bi9021186;
RA Mahlstedt S.A., Walsh C.T.;
RT "Investigation of anticapsin biosynthesis reveals a four-enzyme pathway to
RT tetrahydrotyrosine in Bacillus subtilis.";
RL Biochemistry 49:912-923(2010).
RN [5]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, AND PATHWAY.
RX PubMed=22765234; DOI=10.1021/bi3006362;
RA Parker J.B., Walsh C.T.;
RT "Stereochemical outcome at four stereogenic centers during conversion of
RT prephenate to tetrahydrotyrosine by BacABGF in the bacilysin pathway.";
RL Biochemistry 51:5622-5632(2012).
RN [6]
RP X-RAY CRYSTALLOGRAPHY (1.75 ANGSTROMS) IN COMPLEX WITH NADP, FUNCTION,
RP CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, MUTAGENESIS OF LYS-113;
RP TYR-117; SER-155; ASN-158 AND SER-250, SUBUNIT, AND SUBSTRATE SPECIFICITY.
RX PubMed=23519407; DOI=10.1107/s0907444912046690;
RA Rajavel M., Perinbam K., Gopal B.;
RT "Structural insights into the role of Bacillus subtilis YwfH (BacG) in
RT tetrahydrotyrosine synthesis.";
RL Acta Crystallogr. D 69:324-332(2013).
CC -!- FUNCTION: Along with the bacABCDEF operon, BacG is involved in the
CC biosynthesis of the nonribosomally synthesized dipeptide antibiotic
CC bacilysin, composed of L-alanine and L-anticapsin. Bacilysin is an
CC irreversible inactivator of the glutaminase domain of glucosamine
CC synthetase (PubMed:20052993). BacG catalyzes the stereoselective
CC reduction of exocyclic-delta(3),delta(5)-dihydro-hydroxyphenylpyruvate
CC (ex-H2HPP), adding a pro-S hydride equivalent to C4 position to yield
CC tetrahydro-hydroxyphenylpyruvate (H4HPP) (PubMed:22765234,
CC PubMed:23519407). Although the 3Z,7R-ex-H2HPP isomer is kinetically
CC disfavored by BacB and produced in a smaller quantity than 3E,7R-ex-
CC H2HPP, it is the preferred substrate for the conjugate reduction
CC reaction of BacG (PubMed:22765234, PubMed:23519407).
CC {ECO:0000269|PubMed:20052993, ECO:0000269|PubMed:22765234,
CC ECO:0000269|PubMed:23519407}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=90 uM for 3E,7R-ex-H2HPP {ECO:0000269|PubMed:22765234};
CC KM=178 uM for ex-H2HPP {ECO:0000269|PubMed:23519407};
CC KM=2700 uM for 3Z,7R-ex-H2HPP {ECO:0000269|PubMed:22765234};
CC Vmax=92.82 nmol/sec/ug enzyme {ECO:0000269|PubMed:23519407};
CC Note=kcat is 71.3 min(-1) for reductase activity with 3Z,7R-ex-H2HPP
CC as substrate. kcat is 33.2 min(-1) for reductase activity with 3E,7R-
CC ex-H2HPP as substrate. kcat is 34.38 sec(-1) for reductase activity
CC with ex-H2HPP as substrate. {ECO:0000269|PubMed:23519407};
CC -!- PATHWAY: Antibiotic biosynthesis; bacilysin biosynthesis.
CC {ECO:0000269|PubMed:22765234, ECO:0000305|PubMed:12372825,
CC ECO:0000305|PubMed:20052993}.
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:23519407}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000305}.
CC -!- INDUCTION: The compound guanosine 5'-diphosphate 3'-diphosphate (ppGpp)
CC is essential for the transcription of the bacABCDEF operon and BacG,
CC and GTP regulates the transcription of both this operon and ywfH via
CC the CodY-mediated regulation system. {ECO:0000269|PubMed:12372825}.
CC -!- SIMILARITY: Belongs to the short-chain dehydrogenases/reductases (SDR)
CC family. {ECO:0000305}.
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DR EMBL; X73124; CAA51642.1; -; Genomic_DNA.
DR EMBL; AL009126; CAB15795.1; -; Genomic_DNA.
DR PIR; S39741; S39741.
DR RefSeq; NP_391648.1; NC_000964.3.
DR RefSeq; WP_003244095.1; NZ_JNCM01000034.1.
DR PDB; 3U49; X-ray; 1.75 A; A/B/C/D=1-259.
DR PDB; 3U4C; X-ray; 2.03 A; A=1-259.
DR PDB; 3U4D; X-ray; 2.70 A; A=1-259.
DR PDBsum; 3U49; -.
DR PDBsum; 3U4C; -.
DR PDBsum; 3U4D; -.
DR AlphaFoldDB; P39644; -.
DR SMR; P39644; -.
DR STRING; 224308.BSU37680; -.
DR PaxDb; P39644; -.
DR PRIDE; P39644; -.
DR EnsemblBacteria; CAB15795; CAB15795; BSU_37680.
DR GeneID; 937215; -.
DR KEGG; bsu:BSU37680; -.
DR PATRIC; fig|224308.179.peg.4080; -.
DR eggNOG; COG1028; Bacteria.
DR InParanoid; P39644; -.
DR OMA; RIIMMST; -.
DR PhylomeDB; P39644; -.
DR BioCyc; BSUB:BSU37680-MON; -.
DR BioCyc; MetaCyc:MON-19125; -.
DR BRENDA; 1.3.1.B7; 658.
DR UniPathway; UPA00100; -.
DR Proteomes; UP000001570; Chromosome.
DR GO; GO:0005737; C:cytoplasm; IEA:UniProtKB-SubCell.
DR GO; GO:0050661; F:NADP binding; IDA:UniProtKB.
DR GO; GO:0016628; F:oxidoreductase activity, acting on the CH-CH group of donors, NAD or NADP as acceptor; IDA:UniProtKB.
DR GO; GO:0017000; P:antibiotic biosynthetic process; IDA:UniProtKB.
DR InterPro; IPR036291; NAD(P)-bd_dom_sf.
DR InterPro; IPR002347; SDR_fam.
DR PRINTS; PR00081; GDHRDH.
DR SUPFAM; SSF51735; SSF51735; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Antibiotic biosynthesis; Cytoplasm; NADP; Oxidoreductase;
KW Reference proteome.
FT CHAIN 1..259
FT /note="NADPH-dependent reductase BacG"
FT /id="PRO_0000054851"
FT BINDING 12..15
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:23519407"
FT BINDING 34..36
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:23519407"
FT BINDING 62..63
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:23519407"
FT BINDING 90
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:23519407"
FT BINDING 113
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:23519407"
FT BINDING 185..191
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:23519407"
FT MUTAGEN 113
FT /note="K->A: 5- and 2-fold decrease of the catalytic
FT efficiency and the affinity for ex-H2HPP, respectively."
FT /evidence="ECO:0000269|PubMed:23519407"
FT MUTAGEN 117
FT /note="Y->A: 6- and 3-fold decrease of the catalytic
FT efficiency and the affinity for ex-H2HPP, respectively."
FT /evidence="ECO:0000269|PubMed:23519407"
FT MUTAGEN 155
FT /note="S->A: 5.5- and 3-fold decrease of the catalytic
FT efficiency and the affinity for ex-H2HPP, respectively."
FT /evidence="ECO:0000269|PubMed:23519407"
FT MUTAGEN 158
FT /note="N->A: 5- and 2-fold decrease of the catalytic
FT efficiency and the affinity for ex-H2HPP, respectively."
FT /evidence="ECO:0000269|PubMed:23519407"
FT MUTAGEN 250
FT /note="S->A: 1.5- and 2-fold decrease of the catalytic
FT efficiency and the affinity for ex-H2HPP, respectively."
FT /evidence="ECO:0000269|PubMed:23519407"
FT STRAND 5..10
FT /evidence="ECO:0007829|PDB:3U49"
FT HELIX 14..25
FT /evidence="ECO:0007829|PDB:3U49"
FT STRAND 29..32
FT /evidence="ECO:0007829|PDB:3U49"
FT HELIX 37..50
FT /evidence="ECO:0007829|PDB:3U49"
FT STRAND 56..58
FT /evidence="ECO:0007829|PDB:3U49"
FT HELIX 66..80
FT /evidence="ECO:0007829|PDB:3U49"
FT STRAND 85..88
FT /evidence="ECO:0007829|PDB:3U49"
FT TURN 98..100
FT /evidence="ECO:0007829|PDB:3U49"
FT HELIX 103..112
FT /evidence="ECO:0007829|PDB:3U49"
FT HELIX 114..129
FT /evidence="ECO:0007829|PDB:3U49"
FT TURN 130..132
FT /evidence="ECO:0007829|PDB:3U49"
FT STRAND 134..139
FT /evidence="ECO:0007829|PDB:3U49"
FT STRAND 144..146
FT /evidence="ECO:0007829|PDB:3U49"
FT HELIX 147..150
FT /evidence="ECO:0007829|PDB:3U49"
FT HELIX 152..172
FT /evidence="ECO:0007829|PDB:3U49"
FT HELIX 173..175
FT /evidence="ECO:0007829|PDB:3U49"
FT STRAND 177..183
FT /evidence="ECO:0007829|PDB:3U49"
FT HELIX 191..205
FT /evidence="ECO:0007829|PDB:3U49"
FT HELIX 210..216
FT /evidence="ECO:0007829|PDB:3U4C"
FT HELIX 226..237
FT /evidence="ECO:0007829|PDB:3U49"
FT HELIX 239..241
FT /evidence="ECO:0007829|PDB:3U49"
FT STRAND 248..250
FT /evidence="ECO:0007829|PDB:3U49"
SQ SEQUENCE 259 AA; 28022 MW; 212075C4E637F8B4 CRC64;
MSKRTAFVMG ASQGIGKAIA LKLADQHFSL VINSRNLDNI ESVKEDILAK HPEASVIVLA
GDMSDQHTRA GIFQKIESQC GRLDVLINNI PGGAPDTFDN CNIEDMTATF TQKTVAYIDA
IKRASSLMKQ NEFGRIINIV GNLWKEPGAN MFTNSMMNAA LINASKNISI QLAPHNITVN
CLNPGFIATD RYHQFVENVM KKNSISKQKA EEQIASGIPM KRVGSAEETA ALAAFLASEE
ASYITGQQIS ADGGSMKSI
