BRENDA - Enzyme Database show
show all sequences of 2.1.1.182

Control of substrate specificity by a single active site residue of the KsgA methyltransferase

O'Farrell, H.C.; Musayev, F.N.; Scarsdale, J.N.; Rife, J.P.; Biochemistry 51, 466-474 (2012)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
expression of wild-type and mutant enzymes in Escherichia coli strain XL1-Blue
Escherichia coli
Engineering
Amino acid exchange
Commentary
Organism
C168A
site-directed mutagenesis, the mutation does not affect KsgA activity
Escherichia coli
L114P
site-directed mutagenesis of the active site residue, the KsgA mutant shows diminished overall activity, and impaired ability to methylate the N6-methyladenosine intermediate to produce N6,N6-dimethyladenosine. Reduced activity is not due to disruption of 30S substrate binding
Escherichia coli
N113A
site-directed mutagenesis of the active site residue, the KsgA mutant shows diminishes activity to a level comparable to L114P without affecting the methylation of N6-methyladenosine. Reduced activity is not due to disruption of 30S substrate binding
Escherichia coli
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
4 S-adenosyl-L-methionine + adenine1518/adenine1519 in 16S rRNA
Escherichia coli
-
4 S-adenosyl-L-homocysteine + N6-dimethyladenine1518/N6-dimethyladenine1519 in 16S rRNA
-
-
?
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Escherichia coli
-
-
-
Purification (Commentary)
Commentary
Organism
recombinant wild-type and mutant enzymes from Escherichia coli strain XL1-Blue
Escherichia coli
Reaction
Reaction
Commentary
Organism
4 S-adenosyl-L-methionine + adenine1518/adenine1519 in 16S rRNA = 4 S-adenosyl-L-homocysteine + N6-dimethyladenine1518/N6-dimethyladenine1519 in 16S rRNA
reaction mechanism, overview
Escherichia coli
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
4 S-adenosyl-L-methionine + adenine1518/adenine1519 in 16S rRNA
-
718932
Escherichia coli
4 S-adenosyl-L-homocysteine + N6-dimethyladenine1518/N6-dimethyladenine1519 in 16S rRNA
-
-
-
?
4 S-adenosyl-L-methionine + adenine1518/adenine1519 in 16S rRNA
binding of 30S subunit and S-adenosyl-L-methionine, structure, overview. After the S-adenosyl-L-methionine addition, KsgA dissociates rapidly from the subunits. The binding of KsgA to substrate is complex and requires regions of rRNA well beyond helix 45, including regions of the 790 loop
718932
Escherichia coli
4 S-adenosyl-L-homocysteine + N6-dimethyladenine1518/N6-dimethyladenine1519 in 16S rRNA
-
-
-
?
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
30
-
assay at
Escherichia coli
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.4
-
assay at
Escherichia coli
Cofactor
Cofactor
Commentary
Organism
Structure
S-adenosyl-L-methionine
-
Escherichia coli
IC50 Value
IC50 Value
IC50 Value Maximum
Commentary
Organism
Inhibitor
Structure
additional information
-
IC50 values of unlabeled wild-type or mutant protein competing with fluorescently labeled wild-type protein for binding to 30S subunits
Escherichia coli
additional information
Cloned(Commentary) (protein specific)
Commentary
Organism
expression of wild-type and mutant enzymes in Escherichia coli strain XL1-Blue
Escherichia coli
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
S-adenosyl-L-methionine
-
Escherichia coli
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
C168A
site-directed mutagenesis, the mutation does not affect KsgA activity
Escherichia coli
L114P
site-directed mutagenesis of the active site residue, the KsgA mutant shows diminished overall activity, and impaired ability to methylate the N6-methyladenosine intermediate to produce N6,N6-dimethyladenosine. Reduced activity is not due to disruption of 30S substrate binding
Escherichia coli
N113A
site-directed mutagenesis of the active site residue, the KsgA mutant shows diminishes activity to a level comparable to L114P without affecting the methylation of N6-methyladenosine. Reduced activity is not due to disruption of 30S substrate binding
Escherichia coli
IC50 Value (protein specific)
IC50 Value
IC50 Value Maximum
Commentary
Organism
Inhibitor
Structure
additional information
-
IC50 values of unlabeled wild-type or mutant protein competing with fluorescently labeled wild-type protein for binding to 30S subunits
Escherichia coli
additional information
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
4 S-adenosyl-L-methionine + adenine1518/adenine1519 in 16S rRNA
Escherichia coli
-
4 S-adenosyl-L-homocysteine + N6-dimethyladenine1518/N6-dimethyladenine1519 in 16S rRNA
-
-
?
Purification (Commentary) (protein specific)
Commentary
Organism
recombinant wild-type and mutant enzymes from Escherichia coli strain XL1-Blue
Escherichia coli
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
4 S-adenosyl-L-methionine + adenine1518/adenine1519 in 16S rRNA
-
718932
Escherichia coli
4 S-adenosyl-L-homocysteine + N6-dimethyladenine1518/N6-dimethyladenine1519 in 16S rRNA
-
-
-
?
4 S-adenosyl-L-methionine + adenine1518/adenine1519 in 16S rRNA
binding of 30S subunit and S-adenosyl-L-methionine, structure, overview. After the S-adenosyl-L-methionine addition, KsgA dissociates rapidly from the subunits. The binding of KsgA to substrate is complex and requires regions of rRNA well beyond helix 45, including regions of the 790 loop
718932
Escherichia coli
4 S-adenosyl-L-homocysteine + N6-dimethyladenine1518/N6-dimethyladenine1519 in 16S rRNA
-
-
-
?
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
30
-
assay at
Escherichia coli
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.4
-
assay at
Escherichia coli
General Information
General Information
Commentary
Organism
evolution
the KsgA family belongs tothe group of S-adenosyl-L-methionine-dependent methyltransferases, known as class I MTases, KsgA is related to DNA adenosine methyltransferases, which transfer only a single methyl group to their target adenosine residue. Part of the discrimination between mono- and dimethyltransferase activity lies in a single residue in the active site, L114; this residue is part of a conserved motif, known as motif IV, which is common to a large group of S-adenosyl-L-methionine-dependent methyltransferases
Escherichia coli
physiological function
the KsgA methyltransferase is universally conserved and plays a key role in regulating ribosome biogenesis. KsgA has a complex reaction mechanism, transferring a total of four methyl groups onto two separate adenosine residues, A1518 and A1519, in the small subunit rRNA. This means that the active site pocket must accept both adenosine and N6-methyladenosine as substrates to catalyze formation of the final product N6,N6-dimethyladenosine
Escherichia coli
General Information (protein specific)
General Information
Commentary
Organism
evolution
the KsgA family belongs tothe group of S-adenosyl-L-methionine-dependent methyltransferases, known as class I MTases, KsgA is related to DNA adenosine methyltransferases, which transfer only a single methyl group to their target adenosine residue. Part of the discrimination between mono- and dimethyltransferase activity lies in a single residue in the active site, L114; this residue is part of a conserved motif, known as motif IV, which is common to a large group of S-adenosyl-L-methionine-dependent methyltransferases
Escherichia coli
physiological function
the KsgA methyltransferase is universally conserved and plays a key role in regulating ribosome biogenesis. KsgA has a complex reaction mechanism, transferring a total of four methyl groups onto two separate adenosine residues, A1518 and A1519, in the small subunit rRNA. This means that the active site pocket must accept both adenosine and N6-methyladenosine as substrates to catalyze formation of the final product N6,N6-dimethyladenosine
Escherichia coli
Other publictions for EC 2.1.1.182
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
733426
Kyuma
16S rRNA methyltransferase Ksg ...
Staphylococcus aureus, Staphylococcus aureus RN4220
Biochimie
119
166-174
2015
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2
2
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733980
Phunpruch
A role for 16S rRNA dimethyltr ...
Mycobacterium tuberculosis, Mycobacterium tuberculosis H37Rv
Int. J. Antimicrob. Agents
41
548-551
2013
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2
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163
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1
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718932
O'Farrell
Control of substrate specifici ...
Escherichia coli
Biochemistry
51
466-474
2012
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1
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3
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2
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1
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1
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2
2
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701730
Ochi
Inactivation of KsgA, a 16S rR ...
Bacillus subtilis
Antimicrob. Agents Chemother.
53
193-201
2009
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1
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702903
Binet
The chlamydial functional homo ...
Chlamydia psittaci, Chlamydia psittaci 6BC, Chlamydia trachomatis, Chlamydia trachomatis L2
BMC Microbiol.
9
0000
2009
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2
2
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704036
Duffin
ksgA mutations confer resistan ...
Neisseria gonorrhoeae
Int. J. Antimicrob. Agents
33
321-327
2009
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1
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1
1
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705141
Demirci
Structural rearrangements in t ...
Thermus thermophilus, Thermus thermophilus HB8 / ATCC 27634 / DSM 579
J. Mol. Biol.
388
271-282
2009
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1
1
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2
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1
1
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705968
Zhang-Akiyama
KsgA, a 16S rRNA adenine methy ...
Escherichia coli
Nucleic Acids Res.
37
2116-2125
2009
-
-
1
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1
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1
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706787
Tu
Structural basis for binding o ...
Aquifex aeolicus
Structure
17
374-385
2009
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705774
Connolly
Mechanistic insight into the r ...
Escherichia coli
Mol. Microbiol.
70
1062-1075
2008
-
1
1
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1
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2
2
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704274
Inoue
Dissection of 16S rRNA methylt ...
Escherichia coli
J. Bacteriol.
189
8510-8518
2007
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4
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1
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701901
Desai
The adenosine dimethyltransfer ...
Escherichia coli
Arch. Biochem. Biophys.
449
57-63
2006
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1
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706715
O'Farrell
Recognition of a complex subst ...
Escherichia coli
RNA
12
725-733
2006
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662620
O'Farrell
Crystal structure of KsgA, a u ...
Escherichia coli
J. Mol. Biol.
339
337-353
2004
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485442
Formenoy
Methylation of the conserved A ...
Escherichia coli
Biochimie
76
1123-1128
1994
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705108
Lafontaine
The DIM1 gene responsible for ...
Escherichia coli
J. Mol. Biol.
241
492-497
1994
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702395
Cunningham
Site-specific mutation of the ...
Escherichia coli
Biochim. Biophys. Acta
1050
18-26
1990
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703844
van Buul
Nucleotide sequence of the ksg ...
Escherichia coli
Gene
38
65-72
1985
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1
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705740
Andresson
Some properties of the ribosom ...
Escherichia coli
Mol. Gen. Genet.
179
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1972
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