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Literature summary 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

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.2.1.103 expressed in Escherichia coli BL21-CodonPlus (DE3)-RIL cells Thermus thermophilus
1.2.1.106
-
 Thermus thermophilus

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.2.1.103 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
1.2.1.106 structures of LysY complexed with amino group-carrier protein LysW-gamma-aminoadipic semialdehyde and LysW-gamma-alpha-aminoadipate, respectively. In both structures, the globular domain of LysW is recognized by positively charged residues on helix 9 and the beta11-alpha10 loop of LysY through conformational changes Thermus thermophilus

Protein Variants

EC Number Protein Variants Comment Organism
1.2.1.103 C148A inactive Thermus thermophilus
1.2.1.103 H209A the mutant exhibits similar activity to that of the wildtype enzyme Thermus thermophilus
1.2.1.103 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
1.2.1.103 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
1.2.1.103 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
1.2.1.103 R278A inactive Thermus thermophilus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.2.1.103 0.0078
-
 [amino group carrier protein LysW]-gamma-aminoadipic semialdehyde mutant enzyme H209A, at pH 7.5 and 60°C Thermus thermophilus
1.2.1.103 0.0098
-
 [amino group carrier protein LysW]-gamma-aminoadipic semialdehyde wild type enzyme, at pH 9.5 and 60°C Thermus thermophilus
1.2.1.103 0.011
-
 [amino group carrier protein LysW]-gamma-aminoadipic semialdehyde mutant enzyme H209A, at pH 9.5 and 60°C Thermus thermophilus
1.2.1.103 0.011
-
 [amino group carrier protein LysW]-gamma-aminoadipic semialdehyde wild type enzyme, at pH 7.5 and 60°C Thermus thermophilus
1.2.1.103 0.014
-
 NADP+ wild type enzyme, at pH 9.5 and 60°C Thermus thermophilus
1.2.1.103 0.037
-
 [amino group carrier protein LysW]-gamma-aminoadipic semialdehyde mutant enzyme R102A, at pH 9.5 and 60°C Thermus thermophilus
1.2.1.103 0.17
-
 [amino group carrier protein LysW]-gamma-aminoadipic semialdehyde mutant enzyme R258A, at pH 9.5 and 60°C Thermus thermophilus
1.2.1.103 0.39
-
 [amino group carrier protein LysW]-gamma-aminoadipic semialdehyde mutant enzyme R195A, at pH 9.5 and 60°C Thermus thermophilus
1.2.1.103 0.69
-
 [amino group carrier protein LysW]-gamma-aminoadipic semialdehyde mutant enzyme K271A, at pH 9.5 and 60°C Thermus thermophilus
1.2.1.103 12
-
 phosphate wild type enzyme, at pH 9.5 and 60°C Thermus thermophilus
1.2.1.106 0.0098
-
 [amino-group carrier protein LysW]-C-terminal-gamma-aminoadipic semialdehyde pH 9.5, 60°C Thermus thermophilus
1.2.1.106 0.014
-
 NADP+ pH 9.5, 60°C Thermus thermophilus
1.2.1.106 12
-
 phosphate pH 9.5, 60°C Thermus thermophilus

Natural Substrates/ Products (Substrates)

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

Organism

EC Number Organism UniProt Comment Textmining
1.2.1.103 Thermus thermophilus O50146
-
-
1.2.1.106 Thermus thermophilus O50146 cf. EC 1.2.1.103
-
2.7.2.19 Thermus thermophilus O50147
-
-

Purification (Commentary)

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

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.2.1.106 10.1
-
 NADPH oxidation, pH 8, 60°C Thermus thermophilus

Substrates and Products (Substrate)

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

Subunits

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

Synonyms

EC Number Synonyms Comment Organism
1.2.1.103 lysY
-
 Thermus thermophilus
1.2.1.106 lysY
-
 Thermus thermophilus
2.7.2.19 lysZ
-
 Thermus thermophilus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.2.1.103 0.0063
-
 [amino group carrier protein LysW]-gamma-aminoadipic semialdehyde mutant enzyme R102A, at pH 9.5 and 60°C Thermus thermophilus
1.2.1.103 0.0082
-
 [amino group carrier protein LysW]-gamma-aminoadipic semialdehyde mutant enzyme H209A, at pH 7.5 and 60°C Thermus thermophilus
1.2.1.103 0.011
-
 [amino group carrier protein LysW]-gamma-aminoadipic semialdehyde wild type enzyme, at pH 7.5 and 60°C Thermus thermophilus
1.2.1.103 0.19
-
 [amino group carrier protein LysW]-gamma-aminoadipic semialdehyde mutant enzyme R195A, at pH 9.5 and 60°C Thermus thermophilus
1.2.1.103 0.21
-
 [amino group carrier protein LysW]-gamma-aminoadipic semialdehyde mutant enzyme K271A, at pH 9.5 and 60°C Thermus thermophilus
1.2.1.103 0.5
-
 [amino group carrier protein LysW]-gamma-aminoadipic semialdehyde mutant enzyme H209A, at pH 9.5 and 60°C Thermus thermophilus
1.2.1.103 0.66
-
 [amino group carrier protein LysW]-gamma-aminoadipic semialdehyde mutant enzyme R258A, at pH 9.5 and 60°C Thermus thermophilus
1.2.1.103 0.96
-
 [amino group carrier protein LysW]-gamma-aminoadipic semialdehyde wild type enzyme, at pH 9.5 and 60°C Thermus thermophilus
1.2.1.103 1.1
-
 phosphate wild type enzyme, at pH 9.5 and 60°C Thermus thermophilus
1.2.1.103 1.2
-
 NADP+ wild type enzyme, at pH 9.5 and 60°C Thermus thermophilus
1.2.1.106 0.96
-
 [amino-group carrier protein LysW]-C-terminal-gamma-aminoadipic semialdehyde pH 9.5, 60°C Thermus thermophilus
1.2.1.106 1.1
-
 phosphate pH 9.5, 60°C Thermus thermophilus
1.2.1.106 1.2
-
 NADP+ pH 9.5, 60°C Thermus thermophilus

Cofactor

EC Number Cofactor Comment Organism Structure
1.2.1.103 NADP+
-
 Thermus thermophilus
1.2.1.103 NADPH
-
 Thermus thermophilus
1.2.1.106 additional information no cofactor: NADH Thermus thermophilus
1.2.1.106 NADP+
-
 Thermus thermophilus
1.2.1.106 NADPH
-
 Thermus thermophilus

General Information

EC Number General Information Comment Organism
1.2.1.106 metabolism for lysine biosynthesis, amino group-carrier protein LysW-gamma-aminoadipic acid is phosphorylated by LysZ to form LysW-gamma-aminoadipic acid phosphate, which is subsequently reduced to LysW-gamma-aminoadipic semialdehyde through a reaction catalyzed by LysY Thermus thermophilus
2.7.2.19 physiological function the enzyme participates in an L-arginine biosynthetic pathway Thermus thermophilus

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.2.1.103 0.0089
-
 phosphate wild type enzyme, at pH 9.5 and 60°C Thermus thermophilus
1.2.1.103 0.17
-
 [amino group carrier protein LysW]-gamma-aminoadipic semialdehyde mutant enzyme R102A, at pH 9.5 and 60°C Thermus thermophilus
1.2.1.103 0.31
-
 [amino group carrier protein LysW]-gamma-aminoadipic semialdehyde mutant enzyme K271A, at pH 9.5 and 60°C Thermus thermophilus
1.2.1.103 0.49
-
 [amino group carrier protein LysW]-gamma-aminoadipic semialdehyde mutant enzyme R195A, at pH 9.5 and 60°C Thermus thermophilus
1.2.1.103 1.1
-
 [amino group carrier protein LysW]-gamma-aminoadipic semialdehyde mutant enzyme H209A, at pH 7.5 and 60°C Thermus thermophilus
1.2.1.103 4
-
 [amino group carrier protein LysW]-gamma-aminoadipic semialdehyde mutant enzyme R258A, at pH 9.5 and 60°C Thermus thermophilus
1.2.1.103 15
-
 [amino group carrier protein LysW]-gamma-aminoadipic semialdehyde wild type enzyme, at pH 7.5 and 60°C Thermus thermophilus
1.2.1.103 47
-
 [amino group carrier protein LysW]-gamma-aminoadipic semialdehyde mutant enzyme H209A, at pH 9.5 and 60°C Thermus thermophilus
1.2.1.103 81
-
 NADP+ wild type enzyme, at pH 9.5 and 60°C Thermus thermophilus
1.2.1.103 98
-
 [amino group carrier protein LysW]-gamma-aminoadipic semialdehyde wild type enzyme, at pH 9.5 and 60°C Thermus thermophilus
1.2.1.106 0.0089
-
 phosphate pH 9.5, 60°C Thermus thermophilus
1.2.1.106 81
-
 NADP+ pH 9.5, 60°C Thermus thermophilus
1.2.1.106 98
-
 [amino-group carrier protein LysW]-C-terminal-gamma-aminoadipic semialdehyde pH 9.5, 60°C Thermus thermophilus