Literature summary for 6.3.4.19 extracted from

Nakanishi, K.; Fukai, S.; Ikeuchi, Y.; Soma, A.; Sekine, Y.; Suzuki, T.; Nureki, O.
Structural basis for lysidine formation by ATP pyrophosphatase accompanied by a lysine-specific loop and a tRNA-recognition domain. (2005), Proc. Natl. Acad. Sci. USA, 102, 7487-7492.

Crystallization (Commentary)

Crystallization (Comment)	Organism
crystal structure of TilS at 2.42 A resolution. Structural and functional comparisons with Escherichia coli TilS reveals that the two TilS enzymes discriminate premodified tRNAIle2 from premodified tRNAMet by strategies similar to that used by IleRS, but in distinct manners	Aquifex aeolicus
structural and functional comparisons of Escherichia coli TilS and Axifex aeolicus TilS reveal that the two TilS enzymes discriminate premodified tRNAIle2 from premodified tRNAMet bystrategies similar to that used by IleRS, but in distinct manners	Escherichia coli

Protein Variants

Protein Variants	Comment	Organism
D137A	no activity detectable	Aquifex aeolicus
D191A	no activity detectable	Aquifex aeolicus
D36A	no activity detectable	Aquifex aeolicus
E140A	Km-value for L-lysine is 2.2fold higher than wild-type value, Km-value for ATP is about 7fold higher than wild-type value, Km-value for [tRNAIle2]-cytidine34 is 4.7fold higher than wild-type value	Aquifex aeolicus
H133A	Km-value for L-lysine is 1.6 fold higher than wild-type value, Km-value for ATP is 2.5fold lower than wild-type value, Km-value for [tRNAIle2]-cytidine34 is similar to wild-type value	Aquifex aeolicus
N194A	no activity detectable	Aquifex aeolicus
R113A	no activity detectable	Aquifex aeolicus
R174A	Km-value for L-lysine is 1.8fold higher than wild-type value, Km-value for ATP is 2.1fold higher than wild-type value, Km-value for [tRNAIle2]-cytidine34 is 6fold higher than wild-type value	Aquifex aeolicus
R201A	no activity detectable	Aquifex aeolicus
R205A	Km-value for L-lysine is 1.6fold higher than wild-type value, Km-value for ATP is 14.8fold higher than wild-type value, Km-value for [tRNAIle2]-cytidine34 is 3.6fold higher than wild-type value	Aquifex aeolicus
S37A	Km-value for ATP is 13fold higher than wild-type value	Aquifex aeolicus
W188A	Km-value for ATP is 5fold higher than wild-type value	Aquifex aeolicus
Y114A	Km-value for L-lysine is about 5fold higher than wild-type value, Km-value for ATP is about 8fold higher than wild-type value, Km-value for [tRNAIle2]-cytidine34 is 2.2fold higher than wild-type value	Aquifex aeolicus

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
0.0019	-	[tRNAIle2]-cytidine34	pH 7.8, 60°C, mutant enzyme E106A	Aquifex aeolicus
0.0033	-	[tRNAIle2]-cytidine34	pH 7.8, 60°C,wild-type enzyme	Aquifex aeolicus
0.0035	-	[tRNAIle2]-cytidine34	pH 7.8, 60°C, mutant enzyme H133A	Aquifex aeolicus
0.0074	-	[tRNAIle2]-cytidine34	pH 7.8, 60°C, mutant enzyme Y114A	Aquifex aeolicus
0.0077	-	ATP	pH 7.8, 60°C, mutant enzyme H133A	Aquifex aeolicus
0.0119	-	[tRNAIle2]-cytidine34	pH 7.8, 60°C, mutant enzyme R205A	Aquifex aeolicus
0.0157	-	[tRNAIle2]-cytidine34	pH 7.8, 60°C, mutant enzyme E140A	Aquifex aeolicus
0.019	-	ATP	pH 7.8, 60°C, wild-type enzyme	Aquifex aeolicus
0.0202	-	[tRNAIle2]-cytidine34	pH 7.8, 60°C, mutant enzyme R174A	Aquifex aeolicus
0.0385	-	ATP	pH 7.8, 60°C, mutant enzyme E106A	Aquifex aeolicus
0.0412	-	ATP	pH 7.8, 60°C, mutant enzyme R174A	Aquifex aeolicus
0.0412	-	ATP	pH 7.8, 60°C, mutant enzyme W188A	Aquifex aeolicus
0.135	-	ATP	pH 7.8, 60°C, mutant enzyme E140A	Aquifex aeolicus
0.155	-	ATP	pH 7.8, 60°C, mutant enzyme Y114A	Aquifex aeolicus
0.255	-	ATP	pH 7.8, 60°C, mutant enzyme S37A	Aquifex aeolicus
0.287	-	ATP	pH 7.8, 60°C, mutant enzyme R205A	Aquifex aeolicus
0.629	-	L-lysine	pH 7.8, 60°C, wild-type enzyme	Aquifex aeolicus
1.01	-	L-lysine	pH 7.8, 60°C, mutant enzyme R205A	Aquifex aeolicus
1.04	-	L-lysine	pH 7.8, 60°C, mutant enzyme H133A	Aquifex aeolicus
1.15	-	L-lysine	pH 7.8, 60°C, mutant enzyme R174A	Aquifex aeolicus
1.3	-	L-lysine	pH 7.8, 60°C, mutant enzyme E106A	Aquifex aeolicus
1.37	-	L-lysine	pH 7.8, 60°C, mutant enzyme E140A	Aquifex aeolicus
3.05	-	L-lysine	pH 7.8, 60°C, mutant enzyme Y114A	Aquifex aeolicus

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
[tRNAIle2]-cytidine34 + L-lysine + ATP	Escherichia coli	-	[tRNAIle2]-2-L-lysylcytidine34 + AMP + diphosphate	-	?
[tRNAIle2]-cytidine34 + L-lysine + ATP	Aquifex aeolicus	-	[tRNAIle2]-2-L-lysylcytidine34 + AMP + diphosphate	-	?

Organism

Organism	UniProt	Comment	Textmining
Aquifex aeolicus	O67728	-	-
Escherichia coli	-	-	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
[tRNAIle2]-cytidine34 + L-lysine + ATP	-	Escherichia coli	[tRNAIle2]-2-L-lysylcytidine34 + AMP + diphosphate	-	?
[tRNAIle2]-cytidine34 + L-lysine + ATP	-	Aquifex aeolicus	[tRNAIle2]-2-L-lysylcytidine34 + AMP + diphosphate	-	?
[tRNAIle2]-cytidine34 + L-lysine + ATP	C34 of tRNAIle2 is adenylated by an ATP lying across the central hole of the dinucleotide-binding fold domain. A lysine, which is activated at a loop appended to the dinucleotide-binding fold domain, nucleophilically attacks the C2 carbon from the rear	Aquifex aeolicus	[tRNAIle2]-2-L-lysylcytidine34 + AMP + diphosphate	-	?

Subunits

Subunits	Comment	Organism
dimer	-	Escherichia coli
dimer	each subunit consists of the N-terminal dinucleotide-binding fold domain, with a characteristic central hole, and the C-terminal globular domain connected by a long alpha-helical linker	Aquifex aeolicus

Synonyms

Synonyms	Comment	Organism
lysidine synthetase	-	Escherichia coli
lysidine synthetase	-	Aquifex aeolicus
MesJ protein	-	Escherichia coli
TilS	-	Escherichia coli
TilS	-	Aquifex aeolicus

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
additional information	-	additional information	kcat-values for wild-type enzyme and mutant enzymes	Aquifex aeolicus