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Literature summary for 1.2.1.103 extracted from

  • Shimizu, T.; Tomita, T.; Kuzuyama, T.; Nishiyama, M.
    Crystal structure of the LysY.LysW complex from Thermus thermophilus (2016), J. Biol. Chem., 291, 9948-9959 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 1.2.1.103

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21-CodonPlus (DE3)-RIL cells Thermus thermophilus

Crystallization (Commentary)

Crystallization (Comment) Organism
in complex with amino group carrier protein LysW-gamma-alpha-aminoadipate plus NADP+ and amino group carrier protein LysW-gamma-aminoadipic semialdehyde plus NADP+, hanging drop vapor diffusion method, using 20% (w/v) PEG 2000 monomethyl ester and 200 mM calcium acetate Thermus thermophilus

Protein Variants

Protein Variants Comment Organism
C148A inactive Thermus thermophilus
H209A the mutant exhibits similar activity to that of the wildtype enzyme Thermus thermophilus
K271A the mutation results in a 4.6fold decrease in kcat and a 70fold increase in Km value compared to the wild type enzyme Thermus thermophilus
R195A the mutation results in a 5fold decrease in kcat and a 40fold increase in Km value compared to the wild type enzyme Thermus thermophilus
R258A the mutation results in a slight decrease in kcat and a 17fold increase in Km value compared to the wild type enzyme Thermus thermophilus
R278A inactive Thermus thermophilus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0078
-
 [amino group carrier protein LysW]-gamma-aminoadipic semialdehyde mutant enzyme H209A, at pH 7.5 and 60°C Thermus thermophilus
0.0098
-
 [amino group carrier protein LysW]-gamma-aminoadipic semialdehyde wild type enzyme, at pH 9.5 and 60°C Thermus thermophilus
0.011
-
 [amino group carrier protein LysW]-gamma-aminoadipic semialdehyde mutant enzyme H209A, at pH 9.5 and 60°C Thermus thermophilus
0.011
-
 [amino group carrier protein LysW]-gamma-aminoadipic semialdehyde wild type enzyme, at pH 7.5 and 60°C Thermus thermophilus
0.014
-
 NADP+ wild type enzyme, at pH 9.5 and 60°C Thermus thermophilus
0.037
-
 [amino group carrier protein LysW]-gamma-aminoadipic semialdehyde mutant enzyme R102A, at pH 9.5 and 60°C Thermus thermophilus
0.17
-
 [amino group carrier protein LysW]-gamma-aminoadipic semialdehyde mutant enzyme R258A, at pH 9.5 and 60°C Thermus thermophilus
0.39
-
 [amino group carrier protein LysW]-gamma-aminoadipic semialdehyde mutant enzyme R195A, at pH 9.5 and 60°C Thermus thermophilus
0.69
-
 [amino group carrier protein LysW]-gamma-aminoadipic semialdehyde mutant enzyme K271A, at pH 9.5 and 60°C Thermus thermophilus
12
-
 phosphate wild type enzyme, at pH 9.5 and 60°C Thermus thermophilus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
[amino group carrier protein LysW]-gamma-alpha-aminoadipate + NADPH + H+ Thermus thermophilus
-
 [amino group carrier protein LysW]-gamma-aminoadipic semialdehyde + phosphate + NADP+
-
 r
[amino group carrier protein LysW]-gamma-aminoadipic semialdehyde + phosphate + NADP+ Thermus thermophilus
-
 [amino group carrier protein LysW]-gamma-alpha-aminoadipate phosphate + NADPH + H+
-
 r

Organism

Organism UniProt Comment Textmining
Thermus thermophilus O50146
-
-

Purification (Commentary)

Purification (Comment) Organism
Blue-Sepharose column chromatography and Superdex S200 gel filtration Thermus thermophilus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
[amino group carrier protein LysW]-gamma-alpha-aminoadipate + NADPH + H+
-
 Thermus thermophilus [amino group carrier protein LysW]-gamma-aminoadipic semialdehyde + phosphate + NADP+
-
 r
[amino group carrier protein LysW]-gamma-aminoadipic semialdehyde + phosphate + NADP+
-
 Thermus thermophilus [amino group carrier protein LysW]-gamma-alpha-aminoadipate phosphate + NADPH + H+
-
 r

Subunits

Subunits Comment Organism
homotetramer 4 * 38000, SDS-PAGE Thermus thermophilus

Synonyms

Synonyms Comment Organism
lysY
-
 Thermus thermophilus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.0063
-
 [amino group carrier protein LysW]-gamma-aminoadipic semialdehyde mutant enzyme R102A, at pH 9.5 and 60°C Thermus thermophilus
0.0082
-
 [amino group carrier protein LysW]-gamma-aminoadipic semialdehyde mutant enzyme H209A, at pH 7.5 and 60°C Thermus thermophilus
0.011
-
 [amino group carrier protein LysW]-gamma-aminoadipic semialdehyde wild type enzyme, at pH 7.5 and 60°C Thermus thermophilus
0.19
-
 [amino group carrier protein LysW]-gamma-aminoadipic semialdehyde mutant enzyme R195A, at pH 9.5 and 60°C Thermus thermophilus
0.21
-
 [amino group carrier protein LysW]-gamma-aminoadipic semialdehyde mutant enzyme K271A, at pH 9.5 and 60°C Thermus thermophilus
0.5
-
 [amino group carrier protein LysW]-gamma-aminoadipic semialdehyde mutant enzyme H209A, at pH 9.5 and 60°C Thermus thermophilus
0.66
-
 [amino group carrier protein LysW]-gamma-aminoadipic semialdehyde mutant enzyme R258A, at pH 9.5 and 60°C Thermus thermophilus
0.96
-
 [amino group carrier protein LysW]-gamma-aminoadipic semialdehyde wild type enzyme, at pH 9.5 and 60°C Thermus thermophilus
1.1
-
 phosphate wild type enzyme, at pH 9.5 and 60°C Thermus thermophilus
1.2
-
 NADP+ wild type enzyme, at pH 9.5 and 60°C Thermus thermophilus

Cofactor

Cofactor Comment Organism Structure
NADP+
-
 Thermus thermophilus
NADPH
-
 Thermus thermophilus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.0089
-
 phosphate wild type enzyme, at pH 9.5 and 60°C Thermus thermophilus
0.17
-
 [amino group carrier protein LysW]-gamma-aminoadipic semialdehyde mutant enzyme R102A, at pH 9.5 and 60°C Thermus thermophilus
0.31
-
 [amino group carrier protein LysW]-gamma-aminoadipic semialdehyde mutant enzyme K271A, at pH 9.5 and 60°C Thermus thermophilus
0.49
-
 [amino group carrier protein LysW]-gamma-aminoadipic semialdehyde mutant enzyme R195A, at pH 9.5 and 60°C Thermus thermophilus
1.1
-
 [amino group carrier protein LysW]-gamma-aminoadipic semialdehyde mutant enzyme H209A, at pH 7.5 and 60°C Thermus thermophilus
4
-
 [amino group carrier protein LysW]-gamma-aminoadipic semialdehyde mutant enzyme R258A, at pH 9.5 and 60°C Thermus thermophilus
15
-
 [amino group carrier protein LysW]-gamma-aminoadipic semialdehyde wild type enzyme, at pH 7.5 and 60°C Thermus thermophilus
47
-
 [amino group carrier protein LysW]-gamma-aminoadipic semialdehyde mutant enzyme H209A, at pH 9.5 and 60°C Thermus thermophilus
81
-
 NADP+ wild type enzyme, at pH 9.5 and 60°C Thermus thermophilus
98
-
 [amino group carrier protein LysW]-gamma-aminoadipic semialdehyde wild type enzyme, at pH 9.5 and 60°C Thermus thermophilus