GNAT2_STRCL
ID GNAT2_STRCL Reviewed; 393 AA.
AC P0DJQ5; Q53940;
DT 05-SEP-2012, integrated into UniProtKB/Swiss-Prot.
DT 05-SEP-2012, sequence version 1.
DT 03-AUG-2022, entry version 41.
DE RecName: Full=Glutamate N-acetyltransferase 2;
DE EC=2.3.1.35;
DE AltName: Full=Ornithine acetyltransferase 2;
DE AltName: Full=Ornithine transacetylase 2;
DE Short=OATase 2;
DE Contains:
DE RecName: Full=Glutamate N-acetyltransferase 2 alpha chain;
DE Contains:
DE RecName: Full=Glutamate N-acetyltransferase 2 beta chain;
GN Name=oat2; Synonyms=dclD;
OS Streptomyces clavuligerus.
OC Bacteria; Actinobacteria; Streptomycetales; Streptomycetaceae;
OC Streptomyces.
OX NCBI_TaxID=1901;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], AND NOMENCLATURE.
RX PubMed=8529893; DOI=10.1016/0378-1119(95)00560-9;
RA Hodgson J.E., Fosberry A.P., Rawlinson N.S., Ross H.N.M., Neal R.J.,
RA Arnell J.C., Earl A.J., Lawlor E.J.;
RT "Clavulanic acid biosynthesis in Streptomyces clavuligerus: gene cloning
RT and characterization.";
RL Gene 166:49-55(1995).
RN [2]
RP PROTEIN SEQUENCE OF 1-10 AND 181-190, FUNCTION AS AN OATASE, CATALYTIC
RP ACTIVITY, MASS SPECTROMETRY, AUTOCATALYTIC CLEAVAGE, BIOPHYSICOCHEMICAL
RP PROPERTIES, SUBSTRATE SPECIFICITY, AND SUBUNIT.
RX PubMed=11985581; DOI=10.1046/j.1432-1033.2002.02853.x;
RA Kershaw N.J., McNaughton H.J., Hewitson K.S., Hernandez H., Griffin J.,
RA Hughes C., Greaves P., Barton B., Robinson C.V., Schofield C.J.;
RT "ORF6 from the clavulanic acid gene cluster of Streptomyces clavuligerus
RT has ornithine acetyltransferase activity.";
RL Eur. J. Biochem. 269:2052-2059(2002).
RN [3]
RP X-RAY CRYSTALLOGRAPHY (2.75 ANGSTROMS), MUTAGENESIS OF THR-181, AND
RP SUBUNIT.
RX PubMed=15352873; DOI=10.1042/bj20040814;
RA Elkins J.M., Kershaw N.J., Schofield C.J.;
RT "X-ray crystal structure of ornithine acetyltransferase from the clavulanic
RT acid biosynthesis gene cluster.";
RL Biochem. J. 385:565-573(2005).
RN [4]
RP X-RAY CRYSTALLOGRAPHY (2.20 ANGSTROMS), AND SUBUNIT.
RA Iqbal A., Clifton I.J., Schofield C.J.;
RT "Structure of a novel n-acyl-enzyme intermediate of an n- terminal
RT nucleophile (ntn) hydrolase, oat2.";
RL Submitted (SEP-2008) to the PDB data bank.
RN [5]
RP X-RAY CRYSTALLOGRAPHY (2.33 ANGSTROMS) OF 8-393 OF ACYL INTERMEDIATE, AND
RP SUBUNIT.
RX PubMed=19105697; DOI=10.1021/ja807215u;
RA Iqbal A., Clifton I.J., Bagonis M., Kershaw N.J., Domene C., Claridge T.D.,
RA Wharton C.W., Schofield C.J.;
RT "Anatomy of a simple acyl intermediate in enzyme catalysis: combined
RT biophysical and modeling studies on ornithine acetyl transferase.";
RL J. Am. Chem. Soc. 131:749-757(2009).
RN [6]
RP X-RAY CRYSTALLOGRAPHY (2.7 ANGSTROMS) OF ACYL INTERMEDIATE AND IN COMPLEX
RP WITH SUBSTRATE, MUTAGENESIS OF THR-148; THR-149; ASP-150 AND LYS-170, AND
RP SUBUNIT.
RX PubMed=21796301; DOI=10.1039/c1ob05554b;
RA Iqbal A., Clifton I.J., Chowdhury R., Ivison D., Domene C., Schofield C.J.;
RT "Structural and biochemical analyses reveal how ornithine acetyl
RT transferase binds acidic and basic amino acid substrates.";
RL Org. Biomol. Chem. 9:6219-6225(2011).
CC -!- FUNCTION: Catalyzes the biosynthesis of ornithine by transacetylation
CC between N(2)-acetylornithine and glutamate. It can also use L-arginine,
CC L-glutamine and L-lysine as acetyl acceptors.
CC {ECO:0000269|PubMed:11985581}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=L-glutamate + N(2)-acetyl-L-ornithine = L-ornithine + N-
CC acetyl-L-glutamate; Xref=Rhea:RHEA:15349, ChEBI:CHEBI:29985,
CC ChEBI:CHEBI:44337, ChEBI:CHEBI:46911, ChEBI:CHEBI:57805; EC=2.3.1.35;
CC Evidence={ECO:0000269|PubMed:11985581};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=3.6 mM for L-N-acetylornithine (at 37 degrees Celsius and pH 8.0)
CC {ECO:0000269|PubMed:11985581};
CC Vmax=87 nmol/min/mg enzyme {ECO:0000269|PubMed:11985581};
CC pH dependence:
CC Optimum pH is between 7.5-8. {ECO:0000269|PubMed:11985581};
CC -!- PATHWAY: Antibiotic biosynthesis; clavulanate biosynthesis.
CC -!- SUBUNIT: Heterotetramer of two alpha and two beta chains.
CC {ECO:0000269|PubMed:11985581, ECO:0000269|PubMed:15352873,
CC ECO:0000269|PubMed:19105697, ECO:0000269|PubMed:21796301,
CC ECO:0000269|Ref.4}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000250}.
CC -!- MASS SPECTROMETRY: [Glutamate N-acetyltransferase 2 alpha chain]:
CC Mass=18816.2; Method=Electrospray;
CC Evidence={ECO:0000269|PubMed:11985581};
CC -!- MASS SPECTROMETRY: [Glutamate N-acetyltransferase 2 beta chain]:
CC Mass=22811.7; Method=Electrospray;
CC Evidence={ECO:0000269|PubMed:11985581};
CC -!- MISCELLANEOUS: The role of this protein is probably directed towards
CC producing increased intracellular concentrations of arginine for clavam
CC biosynthesis, rather than primary metabolism.
CC {ECO:0000305|PubMed:11985581}.
CC -!- SIMILARITY: Belongs to the ArgJ family. {ECO:0000305}.
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DR EMBL; X84101; CAA58906.1; -; Genomic_DNA.
DR PIR; S57671; S57671.
DR RefSeq; WP_003952513.1; NZ_CP027858.1.
DR PDB; 1VZ6; X-ray; 2.75 A; A/B=1-393.
DR PDB; 1VZ7; X-ray; 3.00 A; A/B/C/D=1-393.
DR PDB; 1VZ8; X-ray; 2.75 A; A/B/C/D=1-393.
DR PDB; 2V4I; X-ray; 2.20 A; A/C/E/G=8-180, B/D/F/H=182-393.
DR PDB; 2VZK; X-ray; 2.33 A; A/C/E/G=8-180, B=181-393, D/F/H=182-393.
DR PDB; 2YEP; X-ray; 2.70 A; A/C/E/G=1-180, B/D/H=182-393, F=181-393.
DR PDBsum; 1VZ6; -.
DR PDBsum; 1VZ7; -.
DR PDBsum; 1VZ8; -.
DR PDBsum; 2V4I; -.
DR PDBsum; 2VZK; -.
DR PDBsum; 2YEP; -.
DR AlphaFoldDB; P0DJQ5; -.
DR SMR; P0DJQ5; -.
DR BRENDA; 2.3.1.35; 5988.
DR UniPathway; UPA00112; -.
DR GO; GO:0005737; C:cytoplasm; IEA:UniProtKB-SubCell.
DR GO; GO:0004042; F:acetyl-CoA:L-glutamate N-acetyltransferase activity; IEA:UniProtKB-UniRule.
DR GO; GO:0004358; F:glutamate N-acetyltransferase activity; IEA:UniProtKB-UniRule.
DR GO; GO:0006526; P:arginine biosynthetic process; IEA:UniProtKB-UniRule.
DR GO; GO:0033050; P:clavulanic acid biosynthetic process; IEA:UniProtKB-UniPathway.
DR CDD; cd02152; OAT; 1.
DR Gene3D; 3.10.20.340; -; 1.
DR HAMAP; MF_01106; ArgJ; 1.
DR InterPro; IPR002813; Arg_biosynth_ArgJ.
DR InterPro; IPR016117; ArgJ-like_dom_sf.
DR InterPro; IPR042195; ArgJ_beta_C.
DR PANTHER; PTHR23100; PTHR23100; 1.
DR Pfam; PF01960; ArgJ; 1.
DR SUPFAM; SSF56266; SSF56266; 1.
DR TIGRFAMs; TIGR00120; ArgJ; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Acyltransferase; Autocatalytic cleavage; Cytoplasm;
KW Direct protein sequencing; Transferase.
FT CHAIN 1..180
FT /note="Glutamate N-acetyltransferase 2 alpha chain"
FT /id="PRO_0000002267"
FT CHAIN 181..393
FT /note="Glutamate N-acetyltransferase 2 beta chain"
FT /id="PRO_0000002268"
FT ACT_SITE 181
FT /note="Nucleophile"
FT BINDING 148..149
FT /ligand="substrate"
FT BINDING 170..173
FT /ligand="substrate"
FT BINDING 181
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:21796301"
FT BINDING 260
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:21796301"
FT BINDING 388
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:21796301"
FT BINDING 393
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:21796301"
FT SITE 111
FT /note="Involved in the stabilization of negative charge on
FT the oxyanion by the formation of the oxyanion hole"
FT /evidence="ECO:0000250"
FT SITE 112
FT /note="Involved in the stabilization of negative charge on
FT the oxyanion by the formation of the oxyanion hole"
FT SITE 180..181
FT /note="Cleavage; by autolysis"
FT MUTAGEN 148
FT /note="T->A: No autoproteolysis."
FT /evidence="ECO:0000269|PubMed:21796301"
FT MUTAGEN 149
FT /note="T->A: Results in a 20:80 mixture of
FT unprocessed:processed proteins."
FT /evidence="ECO:0000269|PubMed:21796301"
FT MUTAGEN 150
FT /note="D->G: Results in a 50:50 mixture of
FT unprocessed:processed proteins."
FT /evidence="ECO:0000269|PubMed:21796301"
FT MUTAGEN 170
FT /note="K->A: No autoproteolysis."
FT /evidence="ECO:0000269|PubMed:21796301"
FT MUTAGEN 181
FT /note="T->A: No autoproteolysis; loss of activity."
FT /evidence="ECO:0000269|PubMed:15352873"
FT STRAND 12..17
FT /evidence="ECO:0007829|PDB:2V4I"
FT STRAND 21..24
FT /evidence="ECO:0007829|PDB:2VZK"
FT STRAND 29..36
FT /evidence="ECO:0007829|PDB:2V4I"
FT STRAND 39..44
FT /evidence="ECO:0007829|PDB:2V4I"
FT HELIX 52..60
FT /evidence="ECO:0007829|PDB:2V4I"
FT STRAND 62..65
FT /evidence="ECO:0007829|PDB:2V4I"
FT STRAND 68..74
FT /evidence="ECO:0007829|PDB:2V4I"
FT HELIX 81..98
FT /evidence="ECO:0007829|PDB:2V4I"
FT HELIX 103..105
FT /evidence="ECO:0007829|PDB:2V4I"
FT STRAND 106..112
FT /evidence="ECO:0007829|PDB:2V4I"
FT HELIX 120..127
FT /evidence="ECO:0007829|PDB:2V4I"
FT STRAND 134..137
FT /evidence="ECO:0007829|PDB:2VZK"
FT HELIX 139..146
FT /evidence="ECO:0007829|PDB:2V4I"
FT STRAND 154..160
FT /evidence="ECO:0007829|PDB:2V4I"
FT STRAND 163..170
FT /evidence="ECO:0007829|PDB:2V4I"
FT STRAND 172..174
FT /evidence="ECO:0007829|PDB:2V4I"
FT STRAND 183..188
FT /evidence="ECO:0007829|PDB:2V4I"
FT HELIX 194..207
FT /evidence="ECO:0007829|PDB:2V4I"
FT HELIX 209..211
FT /evidence="ECO:0007829|PDB:2V4I"
FT STRAND 214..216
FT /evidence="ECO:0007829|PDB:2V4I"
FT STRAND 223..228
FT /evidence="ECO:0007829|PDB:2V4I"
FT HELIX 237..257
FT /evidence="ECO:0007829|PDB:2V4I"
FT STRAND 265..275
FT /evidence="ECO:0007829|PDB:2V4I"
FT HELIX 276..287
FT /evidence="ECO:0007829|PDB:2V4I"
FT HELIX 290..297
FT /evidence="ECO:0007829|PDB:2V4I"
FT HELIX 303..311
FT /evidence="ECO:0007829|PDB:2V4I"
FT TURN 312..315
FT /evidence="ECO:0007829|PDB:1VZ7"
FT TURN 321..323
FT /evidence="ECO:0007829|PDB:2V4I"
FT STRAND 325..328
FT /evidence="ECO:0007829|PDB:2V4I"
FT STRAND 331..334
FT /evidence="ECO:0007829|PDB:2V4I"
FT STRAND 338..341
FT /evidence="ECO:0007829|PDB:2VZK"
FT TURN 342..344
FT /evidence="ECO:0007829|PDB:2VZK"
FT HELIX 345..356
FT /evidence="ECO:0007829|PDB:2V4I"
FT STRAND 357..365
FT /evidence="ECO:0007829|PDB:2V4I"
FT STRAND 368..378
FT /evidence="ECO:0007829|PDB:2V4I"
FT HELIX 382..389
FT /evidence="ECO:0007829|PDB:2V4I"
SQ SEQUENCE 393 AA; 41608 MW; 951ECEE6B9664E48 CRC64;
MSDSTPKTPR GFVVHTAPVG LADDGRDDFT VLASTAPATV SAVFTRSRFA GPSVVLCREA
VADGQARGVV VLARNANVAT GLEGEENARE VREAVARALG LPEGEMLIAS TGVIGRQYPM
ESIREHLKTL EWPAGEGGFD RAARAIMTTD TRPKEVRVSV GGATLVGIAK GVGMLEPDMA
TLLTFFATDA RLDPAEQDRL FRRVMDRTFN AVSIDTDTST SDTAVLFANG LAGEVDAGEF
EEALHTAALA LVKDIASDGE GAAKLIEVQV TGARDDAQAK RVGKTVVNSP LVKTAVHGCD
PNWGRVAMAI GKCSDDTDID QERVTIRFGE VEVYPPKARG DQADDALRAA VAEHLRGDEV
VIGIDLAIAD GAFTVYGCDL TEGYVRLNSE YTT
