Information on EC 2.7.7.47 - streptomycin 3''-adenylyltransferase

Word Map on EC 2.7.7.47
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Select one or more organisms in this record:


The expected taxonomic range for this enzyme is: Bacteria, Eukaryota

top print hide
EC NUMBER
COMMENTARY hide
2.7.7.47
-
top print hide
RECOMMENDED NAME
GeneOntology No.
streptomycin 3''-adenylyltransferase
top print hide
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
ATP + streptomycin = diphosphate + 3''-adenylylstreptomycin
show the reaction diagram
also acts on sprctionomycin
-
-
-
top print hide
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
nucleotidyl group transfer
-
-
-
-
top print hide
SYSTEMATIC NAME
IUBMB Comments
ATP:streptomycin 3''-adenylyltransferase
Also acts on spectinomycin.
top
top print hide
CAS REGISTRY NUMBER
COMMENTARY hide
52660-23-8
-
top print hide
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
Marburg168 BD224
-
-
Manually annotated by BRENDA team
Marburg168 BD224
-
-
Manually annotated by BRENDA team
HZ95
SwissProt
Manually annotated by BRENDA team
W4770, aadA nucleotide sequence
SwissProt
Manually annotated by BRENDA team
gene modified crop, GM cotton events MON 531, MON 15985, MON 88913
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
top print hide
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
physiological function
top print hide
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
ATP + bluensomycin
diphosphate + 3''-adenylylbluensomycin
show the reaction diagram
-
-
-
?
ATP + spectinomycin
diphosphate + 9-adenylylspectinomycin
show the reaction diagram
ATP + spectinomycin
diphosphate + adenylylspectinomycin
show the reaction diagram
ATP + streptobiosamine
diphosphate + 3''-adenylylstreptobiosamine
show the reaction diagram
-
degradation product streptomycin
-
?
ATP + streptomycin
diphosphate + 3''-adenylylstreptomycin
show the reaction diagram
ATP + streptomycin
diphosphate + 9-adenylylstreptomycin
show the reaction diagram
ATP + streptomycin
diphosphate + adenylylstreptomycin
show the reaction diagram
ATP + tetracyclin
diphosphate + 3''adenylyltetracyclin
show the reaction diagram
additional information
?
-
top print hide
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
ATP + streptomycin
diphosphate + 3''-adenylylstreptomycin
show the reaction diagram
top print hide
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
-
ADP, AMP, adenosine, adenine other nucleoside triphosphates or other deoxyadenine-containing compounds cannot replace ATP
-
top print hide
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
MgCl2
required, at least 5 mM
additional information
-
Ni2+, Co2+, Ca2+, Zn2+ and Mn2+ cannot replace Mg2+
top print hide
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
9-O-(adenosine-5'-phophoryl)spectinomycin
-
product inhibition of ANT(3'')-Ii domain
AMP-CPP
-
dead-end inhibition of ANT(3'')-Ii domain
clindamycin
residues Phe34, Asp60, Arg63, Gln64, Leu68, Glu87, Thr89, Val90 are responsible for positioning clindamycin into the active site
kanamycin A
neomycin
-
non-competitive versus streptomycin and uncompetitive versus ATP
streptomycin
strong substrate inhibition at 1mM MgCl2
tobramycin
-
non-competitive versus streptomycin and uncompetitive versus ATP
top print hide
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-mercaptoethanol
-
enhances the degree of streptomycin inactivation
Tris-HCl
-
-
top print hide
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0105 - 0.014
ATP
0.0002 - 0.0014
spectinomycin
0.00008 - 0.0033
streptomycin
top print hide
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.41 - 0.61
ATP
0.26 - 0.63
spectinomycin
0.47 - 0.66
streptomycin
top print hide
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
39 - 55
ATP
1300 - 1600
spectinomycin
140 - 370
streptomycin
top print hide
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.076 - 0.093
9-O-(adenosine-5'-phophoryl)spectinomycin
0.017 - 0.021
AMP-CPP
0.004 - 0.01
kanamycin A
top print hide
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.000046
-
streptomycin
1.3
wild-type, pH 7.5, 23°C
top print hide
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.5
-
assay at
7.8
-
assay at
top print hide
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37
-
assay at
additional information
-
assay is performed at room temperature
top print hide
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.2
-
isoelectric focusing
6
-
isoelectric focusing
top print hide
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
resistance factor RE130, extrachromosomal genetic element
Manually annotated by BRENDA team
-
periplasmic space
-
Manually annotated by BRENDA team
top print hide
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
31000
-
deduced from open reading frame
49000
-
x * 49000, SDS-PAGE
top print hide
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
-
x * 49000, SDS-PAGE
monomer
top print hide
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
modeling of enzyme structure
homology modeling of structure, and ternary complex with ATP and streptomycin or spectinomycin
to 2.5 A resolution. AadA consists of a nucleotidyltransferase domain and an alpha-helical bundle domain. AadA crystallizes as a monomer. ATP binding has to occur before binding of the aminoglycoside substrate, and ATP binding repositions the two domains for aminoglycoside binding in the interdomain cleft
top print hide
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
49.5
melting temperature, wild-type
51
melting temperature, C-terminal deletion mutant
top print hide
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
retention of 90% native-like secondary structure at 0.5 M guanidine hydrochloride, loss of 50% and 75% of secondary structure and 35% and 60% of activity at 0.75 M and 1.5 M guanidine hydrochloride, respectively, at 6 M guanidine hydrochloride loss of secondary structure and activity
-
top print hide
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
DEAE anion-exchange column and gentamycin affinity column
-
recombinant protein
top print hide
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
27.8 kb R-plasmid pTET3 gene cassette aadA9, antibiotic resistance region identified in Mycobacterium fortuitum, cloned in Escherichia coli DH5alphaMCR
-
aadA resistence gene transferred vi filter mating from Enterococcus faecalis W4770 to Enterococcus faecalis JH2-2, PCR amplified aadA gene cloned into vector pCRII and transformed into Escherichia coli DH5-alpha
cloned in Escherichia coli BL21
-
expression in Escherichia coli
gene aadA2B isolated from integron InC as a gene cassette, Escherichia coli cells transformed by electroporation
into the vector pGEM-T for sequencing
streptomycin adenylyltransferase gene isolated from unconjugative plasmids of Enterococcus casseliflavus HZ95 is cloned in Escherichia coli DH5alpha
the plastid expression vector pPRV111A carrying the aadA gene encoding aminoglycoside 3''-adenylyltransferase is used for plastid transformation in Solanum melongena
-
thioredoxin-His6-tagged SMATase fusion protein is produced in a bacterial intracellular expression system mainly in a soluble form
-
top print hide
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D182A
mutation reduces the MIC of streptomycin and spectinomycin
D182N
mutation reduces the MIC of streptomycin and spectinomycin
E87A
mutation reduces the MIC of streptomycin and spectinomycin to that of an AadA null strain
E87Q
mutation reduces the MIC of streptomycin and spectinomycin to that of an AadA null strain
K205A
mutation reduces the MIC of streptomycin and spectinomycin
R192A
mutation reduces the MIC of streptomycin and spectinomycin
W112A
mutation reduces the MIC of spectinomycin to that of an AadA null strain and reduces the MIC of streptomycin
W112F
mutation reduces the MIC of spectinomycin to that of an AadA null strain and reduces the MIC of streptomycin
D182A
-
mutation reduces the MIC of streptomycin and spectinomycin
-
D182N
-
mutation reduces the MIC of streptomycin and spectinomycin
-
E87Q
-
mutation reduces the MIC of streptomycin and spectinomycin to that of an AadA null strain
-
K205A
-
mutation reduces the MIC of streptomycin and spectinomycin
-
R192A
-
mutation reduces the MIC of streptomycin and spectinomycin
-
additional information
top print hide
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
agriculture
-
aminoglycoside 3''-adenylyltransferase confers resistance to streptomycin and spectinomycin
analysis
-
the developed multiplex PCR assay is useful in verifying the GM status of a sample irrespective of the crop and GM trait
diagnostics
medicine
top
Show AA Sequence (1208 entries)
Please use the Sequence Search for a specific query.
top
top
LINKS TO OTHER DATABASES (specific for EC-Number 2.7.7.47)
ExplorEnz
ExPASy
KEGG
MetaCyc
SABIO-RK
NCBI: PubMed, Protein, Nucleotide, Structure, Genome, OMIM
IUBMB Enzyme Nomenclature
PROSITE Database of protein families and domains
InterPro (database of protein families, domains and functional sites)