AADK_BACSU
ID AADK_BACSU Reviewed; 284 AA.
AC P17585; A0A6M3ZEA3;
DT 01-AUG-1990, integrated into UniProtKB/Swiss-Prot.
DT 01-AUG-1990, sequence version 1.
DT 03-AUG-2022, entry version 116.
DE RecName: Full=Aminoglycoside 6-adenylyltransferase {ECO:0000303|PubMed:15984036, ECO:0000303|Ref.5};
DE EC=2.7.7.- {ECO:0000269|PubMed:15984036, ECO:0000269|PubMed:17609790, ECO:0000305|Ref.5};
DE AltName: Full=6-O-adenyl-transferase {ECO:0000303|PubMed:15984036};
DE AltName: Full=AAD(6) {ECO:0000303|Ref.5};
DE AltName: Full=ANT(6) {ECO:0000303|PubMed:15984036, ECO:0000303|PubMed:17609790};
DE AltName: Full=Aminoglycoside inactivating enzyme {ECO:0000303|PubMed:17609790};
DE AltName: Full=Sm inactivating enzyme {ECO:0000303|Ref.5};
DE AltName: Full=Streptomycin 6-adenylyltransferase {ECO:0000303|Ref.5};
GN Name=aadK {ECO:0000303|PubMed:17609790, ECO:0000303|PubMed:8293959,
GN ECO:0000312|EMBL:QJP89398.1}; OrderedLocusNames=BSU26790;
GN ORFNames=HIR78_15755 {ECO:0000312|EMBL:QJP89398.1};
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=2550327; DOI=10.1016/0378-1119(89)90241-2;
RA Ohmiya K., Tanaka T., Noguchi N., O'Hara K., Kono M.;
RT "Nucleotide sequence of the chromosomal gene coding for the aminoglycoside
RT 6-adenylyltransferase from Bacillus subtilis Marburg 168.";
RL Gene 78:377-378(1989).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=168;
RX PubMed=9308178; DOI=10.1099/00221287-143-9-2939;
RA Sorokin A., Bolotin A., Purnelle B., Hilbert H., Lauber J.,
RA Duesterhoeft A., Ehrlich S.D.;
RT "Sequence of the Bacillus subtilis genome region in the vicinity of the lev
RT operon reveals two new extracytoplasmic function RNA polymerase sigma
RT factors SigV and SigZ.";
RL Microbiology 143:2939-2943(1997).
RN [3]
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 [4] {ECO:0000312|EMBL:QJP89398.1}
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=168 {ECO:0000312|EMBL:QJP89398.1};
RA Dragos A., Kovacs A.T.;
RT "Phage recombination drives evolution of spore-forming Bacilli.";
RL Submitted (APR-2020) to the EMBL/GenBank/DDBJ databases.
RN [5]
RP FUNCTION IN STREPTOMYCIN ADENYLATION, BIOPHYSICOCHEMICAL PROPERTIES, AND
RP SUBCELLULAR LOCATION.
RC STRAIN=168 / BD224;
RX DOI=10.1111/j.1574-6968.1987.tb02029.x;
RA Kono M., Ohmiya K., Kanda T., Noguchi N., O'Hara K.;
RT "Purification and characterization of chromosomal streptomycin
RT adenylyltransferase from derivatives of Bacillus subtilis Marburg 168.";
RL FEMS Microbiol. Lett. 40:223-228(1987).
RN [6]
RP FUNCTION IN STREPTOMYCIN ADENYLATION, AND REACTION PRODUCT.
RX PubMed=3137862; DOI=10.1128/aac.32.6.949-b;
RA O'Hara K., Ohmiya K., Kono M.;
RT "Structure of adenylylated streptomycin synthesized enzymatically by
RT Bacillus subtilis.";
RL Antimicrob. Agents Chemother. 32:949-950(1988).
RN [7]
RP BIOTECHNOLOGY.
RX DOI=10.1080/00021369.1989.10869694;
RA Noguchi N., Ohmiya K., Tanaka T., O'Hara K., Kono M.;
RT "Expression of the Aminoglycoside 6-Adenylyltransferase Coding Gene from
RT Bacillus suhtilis in Escherichia coli.";
RL Agric. Biol. Chem. 53:2519-2520(1989).
RN [8]
RP GENE MAPPING, AND DISRUPTION PHENOTYPE.
RX PubMed=8293959; DOI=10.1016/0378-1097(93)90140-w;
RA Noguchi N., Sasatsu M., Kono M.;
RT "Genetic mapping in Bacillus subtilis 168 of the aadK gene which encodes
RT aminoglycoside 6-adenylyltransferase.";
RL FEMS Microbiol. Lett. 114:47-52(1993).
RN [9]
RP FUNCTION, CATALYTIC ACTIVITY, SUBSTRATE SPECIFICITY, AND SUBUNIT.
RX PubMed=15984036; DOI=10.1002/chem.200400941;
RA Corzana F., Cuesta I., Bastida A., Hidalgo A., Latorre M., Gonzalez C.,
RA Garcia-Junceda E., Jimenez-Barbero J., Asensio J.L.;
RT "Molecular recognition of aminoglycoside antibiotics by bacterial defence
RT proteins: NMR study of the structural and conformational features of
RT streptomycin inactivation by Bacillus subtilis aminoglycoside-6-adenyl
RT transferase.";
RL Chemistry 11:5102-5113(2005).
RN [10]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, AND
RP BIOTECHNOLOGY.
RX PubMed=17609790; DOI=10.1039/b704785a;
RA Latorre M., Penalver P., Revuelta J., Asensio J.L., Garcia-Junceda E.,
RA Bastida A.;
RT "Rescue of the streptomycin antibiotic activity by using streptidine as a
RT 'decoy acceptor' for the aminoglycoside-inactivating enzyme adenyl
RT transferase.";
RL Chem. Commun. (Camb.) 27:2829-2831(2007).
RN [11] {ECO:0007744|PDB:2PBE}
RP X-RAY CRYSTALLOGRAPHY (2.65 ANGSTROMS) OF 2-284.
RA Tyagi R., Eswaramoorthy S., Burley S.K., Swaminathan S.;
RT "The crystal structure of an aminoglycoside 6-adenyltransferase from
RT Bacillus subtilis.";
RL Submitted (MAR-2007) to the PDB data bank.
CC -!- FUNCTION: Mediates bacterial resistance to streptomycin
CC (PubMed:3137862, PubMed:17609790). Adenylates streptomycin on the O-6
CC residue (PubMed:3137862, PubMed:15984036, PubMed:17609790). Adenylates
CC streptidine on the O-6 residue (PubMed:17609790). Does not act on
CC spectinomycin, neomycin-B or kanamycin (Ref.5, PubMed:15984036).
CC Specific for ATP and GTP nucleotides incorporating a purine ring. No
CC reaction with CTP or UTP (PubMed:15984036).
CC {ECO:0000269|PubMed:15984036, ECO:0000269|PubMed:17609790,
CC ECO:0000269|PubMed:3137862, ECO:0000269|Ref.5}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=ATP + streptomycin = 6-O-adenylylstreptomycin + diphosphate;
CC Xref=Rhea:RHEA:63236, ChEBI:CHEBI:30616, ChEBI:CHEBI:33019,
CC ChEBI:CHEBI:58007, ChEBI:CHEBI:146262;
CC Evidence={ECO:0000269|PubMed:15984036, ECO:0000269|PubMed:17609790,
CC ECO:0000269|Ref.5};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=GTP + streptomycin = 6-O-guanylylstreptomycin + diphosphate;
CC Xref=Rhea:RHEA:69540, ChEBI:CHEBI:33019, ChEBI:CHEBI:37565,
CC ChEBI:CHEBI:58007, ChEBI:CHEBI:184374;
CC Evidence={ECO:0000269|PubMed:15984036};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=ATP + streptidine = 6-O-adenylylstreptidine + diphosphate;
CC Xref=Rhea:RHEA:69544, ChEBI:CHEBI:30616, ChEBI:CHEBI:33019,
CC ChEBI:CHEBI:184375, ChEBI:CHEBI:184376;
CC Evidence={ECO:0000269|PubMed:17609790};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.08 uM for streptomycin {ECO:0000269|Ref.5};
CC KM=0.04 mM for streptomycin (at pH 7.5 and 25 degrees Celsius)
CC {ECO:0000269|PubMed:17609790};
CC KM=0.6 mM for streptidine (at pH 7.5 and 25 degrees Celsius)
CC {ECO:0000269|PubMed:17609790};
CC Vmax=4.00 pmol/min/mg enzyme with streptomycin as substrate
CC {ECO:0000269|Ref.5};
CC Vmax=0.06 umol/min/mg enzyme with streptomycin as substrate (at pH
CC 7.5 and 25 degrees Celsius) {ECO:0000269|PubMed:17609790};
CC Vmax=0.0012 umol/min/mg enzyme with streptidine as substrate (at pH
CC 7.5 and 25 degrees Celsius) {ECO:0000269|PubMed:17609790};
CC pH dependence:
CC Optimum pH is 8.0. {ECO:0000269|Ref.5};
CC Temperature dependence:
CC Optimum temperature is 35 degrees Celsius. {ECO:0000269|Ref.5};
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:15984036}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000305|Ref.5}.
CC -!- DISRUPTION PHENOTYPE: Very slight reduction in streptomycin MIC.
CC {ECO:0000269|PubMed:8293959}.
CC -!- BIOTECHNOLOGY: Could be used as a selectable marker in B.subtilis if
CC strongly expressed, could be used as a selectable marker in E.coli
CC (Ref.7). Streptidine restores the antibiotic effectiveness of
CC streptomycin in bacterial strains in which they both are co-
CC administered together with a simultaneous overexpression of this enzyme
CC (PubMed:17609790). {ECO:0000269|PubMed:17609790, ECO:0000269|Ref.7}.
CC -!- MISCELLANEOUS: Although this enzyme can be purified from B.subtilis,
CC the bacteria is sensitive to streptomycin in vivo. {ECO:0000269|Ref.5}.
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DR EMBL; M26879; AAA22190.1; -; Genomic_DNA.
DR EMBL; U93875; AAB80892.1; -; Genomic_DNA.
DR EMBL; U93876; AAB80893.1; -; Genomic_DNA.
DR EMBL; AL009126; CAB14620.1; -; Genomic_DNA.
DR EMBL; CP052842; QJP89398.1; -; Genomic_DNA.
DR PIR; JU0059; XXBSG.
DR RefSeq; NP_390556.1; NC_000964.3.
DR RefSeq; WP_003229862.1; NZ_JNCM01000036.1.
DR PDB; 2PBE; X-ray; 2.65 A; A=2-284.
DR PDBsum; 2PBE; -.
DR AlphaFoldDB; P17585; -.
DR SMR; P17585; -.
DR STRING; 224308.BSU26790; -.
DR PaxDb; P17585; -.
DR PRIDE; P17585; -.
DR EnsemblBacteria; CAB14620; CAB14620; BSU_26790.
DR GeneID; 938061; -.
DR KEGG; bsu:BSU26790; -.
DR PATRIC; fig|224308.179.peg.2910; -.
DR eggNOG; ENOG502Z7S1; Bacteria.
DR OMA; FDDCCNE; -.
DR BioCyc; BSUB:BSU26790-MON; -.
DR BioCyc; MetaCyc:BSU26790-MON; -.
DR EvolutionaryTrace; P17585; -.
DR Proteomes; UP000001570; Chromosome.
DR GO; GO:0005737; C:cytoplasm; IDA:UniProtKB.
DR GO; GO:0070566; F:adenylyltransferase activity; IDA:UniProtKB.
DR GO; GO:0042803; F:protein homodimerization activity; IDA:UniProtKB.
DR GO; GO:0071239; P:cellular response to streptomycin; IDA:UniProtKB.
DR Gene3D; 3.30.460.10; -; 1.
DR InterPro; IPR007530; Aminoglycoside_adenylylTfrase.
DR InterPro; IPR043519; NT_sf.
DR Pfam; PF04439; Adenyl_transf; 1.
DR PIRSF; PIRSF000812; AAD; 1.
DR SUPFAM; SSF81301; SSF81301; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Cytoplasm; Nucleotidyltransferase; Reference proteome;
KW Transferase.
FT CHAIN 1..284
FT /note="Aminoglycoside 6-adenylyltransferase"
FT /id="PRO_0000064415"
FT HELIX 4..17
FT /evidence="ECO:0007829|PDB:2PBE"
FT STRAND 21..26
FT /evidence="ECO:0007829|PDB:2PBE"
FT STRAND 43..50
FT /evidence="ECO:0007829|PDB:2PBE"
FT HELIX 52..56
FT /evidence="ECO:0007829|PDB:2PBE"
FT HELIX 60..65
FT /evidence="ECO:0007829|PDB:2PBE"
FT STRAND 68..72
FT /evidence="ECO:0007829|PDB:2PBE"
FT STRAND 78..80
FT /evidence="ECO:0007829|PDB:2PBE"
FT STRAND 88..94
FT /evidence="ECO:0007829|PDB:2PBE"
FT STRAND 99..106
FT /evidence="ECO:0007829|PDB:2PBE"
FT HELIX 107..109
FT /evidence="ECO:0007829|PDB:2PBE"
FT HELIX 110..115
FT /evidence="ECO:0007829|PDB:2PBE"
FT STRAND 123..128
FT /evidence="ECO:0007829|PDB:2PBE"
FT HELIX 139..141
FT /evidence="ECO:0007829|PDB:2PBE"
FT HELIX 149..171
FT /evidence="ECO:0007829|PDB:2PBE"
FT HELIX 175..184
FT /evidence="ECO:0007829|PDB:2PBE"
FT HELIX 186..202
FT /evidence="ECO:0007829|PDB:2PBE"
FT STRAND 205..207
FT /evidence="ECO:0007829|PDB:2PBE"
FT HELIX 210..212
FT /evidence="ECO:0007829|PDB:2PBE"
FT HELIX 215..217
FT /evidence="ECO:0007829|PDB:2PBE"
FT HELIX 221..228
FT /evidence="ECO:0007829|PDB:2PBE"
FT HELIX 236..261
FT /evidence="ECO:0007829|PDB:2PBE"
FT HELIX 268..281
FT /evidence="ECO:0007829|PDB:2PBE"
SQ SEQUENCE 284 AA; 33908 MW; 69C970810B914F9E CRC64;
MRSEQEMMDI FLDFALNDER IRLVTLEGSR TNRNIPPDNF QDYDISYFVT DVESFKENDQ
WLEIFGKRIM MQKPEDMELF PPELGNWFSY IILFEDGNKL DLTLIPIREA EDYFANNDGL
VKVLLDKDSF INYKVTPNDR QYWIKRPTAR EFDDCCNEFW MVSTYVVKGL ARNEILFAID
HLNEIVRPNL LRMMAWHIAS QKGYSFSMGK NYKFMKRYLS NKEWEELMST YSVNGYQEMW
KSLFTCYALF RKYSKAVSEG LAYKYPDYDE GITKYTEGIY CSVK
