AADB1_KLEPN
ID AADB1_KLEPN Reviewed; 177 AA.
AC P0AE05; P08880; P10019;
DT 06-DEC-2005, integrated into UniProtKB/Swiss-Prot.
DT 06-DEC-2005, sequence version 1.
DT 03-AUG-2022, entry version 59.
DE RecName: Full=2''-aminoglycoside nucleotidyltransferase {ECO:0000303|PubMed:2850441};
DE EC=2.7.7.46 {ECO:0000269|PubMed:2850441};
DE AltName: Full=AAD(2'');
DE AltName: Full=Aminoglycoside adenyltransferase 2''-Ia {ECO:0000303|PubMed:28734024};
DE Short=ANT(2'')-Ia {ECO:0000303|PubMed:25564464};
DE AltName: Full=Gentamicin 2''-nucleotidyltransferase;
DE AltName: Full=Gentamicin resistance protein;
GN Name=aadB {ECO:0000303|PubMed:2850441};
OS Klebsiella pneumoniae.
OG Plasmid pBP201, and Plasmid IncM pBWH1.
OC Bacteria; Proteobacteria; Gammaproteobacteria; Enterobacterales;
OC Enterobacteriaceae; Klebsiella/Raoultella group; Klebsiella.
OX NCBI_TaxID=573;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=18440; PLASMID=pBP201; TRANSPOSON=Tn4000;
RX PubMed=2850441; DOI=10.1111/j.1365-2958.1988.tb00081.x;
RA Schmidt F.R.J., Nuecken E.J., Henschke R.B.;
RT "Nucleotide sequence analysis of 2''-aminoglycoside nucleotidyl-transferase
RT ANT(2'') from Tn4000: its relationship with AAD(3'') and impact on Tn21
RT evolution.";
RL Mol. Microbiol. 2:709-717(1988).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC PLASMID=IncM pBWH1;
RA Zieg J., Jiang H., McCabe F., O'Brien T.;
RT "Molecular characterization and comparison of the regions encoding
RT gentamicin resistance (aadB) in pBW1 and pLST1000 and their relationship to
RT Tn21 family of transposons and elements.";
RL Submitted (FEB-1992) to the EMBL/GenBank/DDBJ databases.
RN [3] {ECO:0007744|PDB:4WQK, ECO:0007744|PDB:4WQL}
RP X-RAY CRYSTALLOGRAPHY (1.48 ANGSTROMS) IN COMPLEX WITH MAGNESIUM WITH AND
RP WITHOUT ANTIBIOTIC, FUNCTION, CATALYTIC ACTIVITY, ACTIVE SITE, POSSIBLE
RP REACTION MECHANISM, BIOPHYSICOCHEMICAL PROPERTIES, SUBUNIT, DOMAIN, AND
RP MUTAGENESIS OF ARG-40; HIS-42; ASP-46 AND ASP-86.
RX PubMed=25564464; DOI=10.1128/mbio.02180-14;
RA Cox G., Stogios P.J., Savchenko A., Wright G.D.;
RT "Structural and molecular basis for resistance to aminoglycoside
RT antibiotics by the adenylyltransferase ANT(2)-Ia.";
RL MBio 6:0-0(2015).
RN [4] {ECO:0007744|PDB:5KQJ}
RP STRUCTURE BY NMR, FUNCTION, SUBUNIT, DOMAIN, AND MUTAGENESIS OF ASP-44;
RP ILE-129 AND TYR-135.
RX PubMed=28734024; DOI=10.1002/pro.3224;
RA Bacot-Davis V.R., Bassenden A.V., Sprules T., Berghuis A.M.;
RT "Effect of solvent and protein dynamics in ligand recognition and
RT inhibition of aminoglycoside adenyltransferase 2-Ia.";
RL Protein Sci. 26:1852-1863(2017).
CC -!- FUNCTION: Mediates bacterial resistance to kanamycin, gentamicin,
CC dibekacin, sisomicin and tobramycin by adenylating the 2''-hydroxyl
CC group of these antibiotics. {ECO:0000269|PubMed:25564464,
CC ECO:0000269|PubMed:2850441, ECO:0000269|Ref.2}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=nucleoside triphosphate + gentamicin = diphosphate + 2''-
CC nucleotidylgentamicin.; EC=2.7.7.46;
CC Evidence={ECO:0000269|PubMed:2850441, ECO:0000305|PubMed:25564464};
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000269|PubMed:25564464, ECO:0007744|PDB:4WQK,
CC ECO:0007744|PDB:4WQL};
CC Note=Binds 2 Mg(2+). {ECO:0000269|PubMed:25564464};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=23.9 uM for kanamycin B {ECO:0000269|PubMed:25564464};
CC KM=35 uM for ATP {ECO:0000269|PubMed:25564464};
CC Note=kcat is 15.3 sec(-1) for ATP and 5.2 sec(-1) for kanamycin B.
CC {ECO:0000269|PubMed:25564464};
CC -!- SUBUNIT: Monomer. {ECO:0000269|PubMed:25564464,
CC ECO:0000269|PubMed:28734024}.
CC -!- DOMAIN: Has an N- and C-terminal domain; kanamycin binds in the cleft
CC between the 2 domains. {ECO:0000269|PubMed:25564464,
CC ECO:0000269|PubMed:28734024}.
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DR EMBL; X12618; CAA31139.1; -; Genomic_DNA.
DR EMBL; X64368; CAA45721.1; -; Genomic_DNA.
DR PIR; S04170; XNKBGP.
DR RefSeq; WP_000381802.1; NZ_WYAL01000060.1.
DR RefSeq; YP_009022255.1; NC_023898.1.
DR PDB; 4WQK; X-ray; 1.48 A; A=1-177.
DR PDB; 4WQL; X-ray; 1.73 A; A=1-177.
DR PDB; 5KQJ; NMR; -; A=1-177.
DR PDBsum; 4WQK; -.
DR PDBsum; 4WQL; -.
DR PDBsum; 5KQJ; -.
DR AlphaFoldDB; P0AE05; -.
DR SMR; P0AE05; -.
DR BindingDB; P0AE05; -.
DR GeneID; 67369350; -.
DR BRENDA; 2.7.7.46; 2814.
DR PRO; PR:P0AE05; -.
DR GO; GO:0008871; F:aminoglycoside 2''-nucleotidyltransferase activity; IEA:UniProtKB-EC.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0046677; P:response to antibiotic; IEA:UniProtKB-KW.
DR InterPro; IPR019646; Aminoglyc_AdlTrfase.
DR Pfam; PF10706; Aminoglyc_resit; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Antibiotic resistance; Magnesium; Metal-binding;
KW Nucleotidyltransferase; Plasmid; Transferase; Transposable element.
FT CHAIN 1..177
FT /note="2''-aminoglycoside nucleotidyltransferase"
FT /id="PRO_0000068557"
FT REGION 1..92
FT /note="N-terminal domain"
FT /evidence="ECO:0000269|PubMed:25564464,
FT ECO:0000269|PubMed:28734024"
FT REGION 93..177
FT /note="C-terminal domain"
FT /evidence="ECO:0000269|PubMed:25564464,
FT ECO:0000269|PubMed:28734024"
FT ACT_SITE 86
FT /note="Proton acceptor"
FT /evidence="ECO:0000305|PubMed:25564464"
FT BINDING 44
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:25564464,
FT ECO:0007744|PDB:4WQK, ECO:0007744|PDB:4WQL"
FT BINDING 44
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:25564464,
FT ECO:0007744|PDB:4WQK, ECO:0007744|PDB:4WQL"
FT BINDING 46
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:25564464,
FT ECO:0007744|PDB:4WQK, ECO:0007744|PDB:4WQL"
FT BINDING 46
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:25564464,
FT ECO:0007744|PDB:4WQK, ECO:0007744|PDB:4WQL"
FT BINDING 86
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:25564464,
FT ECO:0007744|PDB:4WQK, ECO:0007744|PDB:4WQL"
FT BINDING 100
FT /ligand="kanamycin A"
FT /ligand_id="ChEBI:CHEBI:58214"
FT /evidence="ECO:0000269|PubMed:25564464,
FT ECO:0007744|PDB:4WQL"
FT MUTAGEN 40
FT /note="R->A: Decreased kcat, decreased affinity for ATP."
FT /evidence="ECO:0000269|PubMed:25564464"
FT MUTAGEN 42
FT /note="H->A: Loss of activity."
FT /evidence="ECO:0000269|PubMed:25564464"
FT MUTAGEN 44
FT /note="D->A: 4-fold reduction in tobramycin modification."
FT /evidence="ECO:0000269|PubMed:28734024"
FT MUTAGEN 46
FT /note="D->A: Loss of activity."
FT /evidence="ECO:0000269|PubMed:25564464"
FT MUTAGEN 86
FT /note="D->A: Loss of activity."
FT /evidence="ECO:0000269|PubMed:25564464"
FT MUTAGEN 129
FT /note="I->A: 3-fold reduction in tobramycin modification."
FT /evidence="ECO:0000269|PubMed:28734024"
FT MUTAGEN 135
FT /note="Y->A: Small decrease in tobramycin modification."
FT /evidence="ECO:0000269|PubMed:28734024"
FT HELIX 4..18
FT /evidence="ECO:0007829|PDB:4WQK"
FT HELIX 27..35
FT /evidence="ECO:0007829|PDB:4WQK"
FT STRAND 43..50
FT /evidence="ECO:0007829|PDB:4WQK"
FT HELIX 51..53
FT /evidence="ECO:0007829|PDB:4WQK"
FT HELIX 54..63
FT /evidence="ECO:0007829|PDB:4WQK"
FT STRAND 67..72
FT /evidence="ECO:0007829|PDB:4WQK"
FT STRAND 75..80
FT /evidence="ECO:0007829|PDB:4WQK"
FT STRAND 83..92
FT /evidence="ECO:0007829|PDB:4WQK"
FT STRAND 94..98
FT /evidence="ECO:0007829|PDB:4WQK"
FT STRAND 113..115
FT /evidence="ECO:0007829|PDB:4WQK"
FT STRAND 118..120
FT /evidence="ECO:0007829|PDB:4WQK"
FT HELIX 125..138
FT /evidence="ECO:0007829|PDB:4WQK"
FT HELIX 141..143
FT /evidence="ECO:0007829|PDB:4WQK"
FT HELIX 146..159
FT /evidence="ECO:0007829|PDB:4WQK"
FT HELIX 161..175
FT /evidence="ECO:0007829|PDB:4WQK"
SQ SEQUENCE 177 AA; 19873 MW; 9A084D96AE99F2F1 CRC64;
MDTTQVTLIH KILAAADERN LPLWIGGGWA IDARLGRVTR KHDDIDLTFP GERRGELEAI
VEMLGGRVME ELDYGFLAEI GDELLDCEPA WWADEAYEIA EAPQGSCPEA AEGVIAGRPV
RCNSWEAIIW DYFYYADEVP PVDWPTKHIE SYRLACTSLG AEKVEVLRAA FRSRYAA
