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Literature summary for 3.1.4.1 extracted from

  • Comeaux, E.Q.; Cuya, S.M.; Kojima, K.; Jafari, N.; Wanzeck, K.C.; Mobley, J.A.; Bjornsti, M.A.; van Waardenburg, R.C.
    Tyrosyl-DNA phosphodiesterase I catalytic mutants reveal an alternative nucleophile that can catalyze substrate cleavage (2015), J. Biol. Chem., 290, 6203-6214 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.1.4.1

Protein Variants

Protein Variants Comment Organism
H182A the mutant shows strongly reduced catalytic efficiency compared to the wild type enzyme, potentiates the cytotoxicity of the Hisgab mutants and induces a DNA topoisomerase 1-independent lethal phenotype in combination with the H432N mutant Saccharomyces cerevisiae
H182A/H432N the mutant protein reveals a dramatic (210fold) reduction in catalytic activity compared to the wild type Saccharomyces cerevisiae
H182A/H432R the mutant protein reveals a dramatic (more than 50%) reduction in catalytic activity compared to the wild type Saccharomyces cerevisiae
H182F catalytically inactive Saccharomyces cerevisiae
H432N the mutant shows strongly reduced catalytic efficiency compared to the wild type enzyme Saccharomyces cerevisiae
H432R the mutant shows strongly reduced catalytic efficiency compared to the wild type enzyme Saccharomyces cerevisiae

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0011
-
 (5'-GATCTAAAAGACTT-3')-phosphotyrosine wild type enzyme, at pH 7.5 and 30°C Saccharomyces cerevisiae
0.0026
-
 (5'-GATCTAAAAGACTT-3')-phosphotyrosine mutant enzyme H432N, at pH 7.5 and 30°C Saccharomyces cerevisiae
0.0034
-
 (5'-GATCTAAAAGACTT-3')-phosphotyrosine mutant enzyme H182A, at pH 7.5 and 30°C Saccharomyces cerevisiae
0.0048
-
 (5'-GATCTAAAAGACTT-3')-phosphotyrosine mutant enzyme H432R, at pH 7.5 and 30°C Saccharomyces cerevisiae

Organism

Organism UniProt Comment Textmining
Saccharomyces cerevisiae P38319
-
-

Purification (Commentary)

Purification (Comment) Organism
SP-Sepharose column chromatography and Flag M2 affinity matrix chromatography Saccharomyces cerevisiae

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(5'-GATCTAAAAGACTT-3')-phosphotyrosine + H2O
-
 Saccharomyces cerevisiae (5'-GATCTAAAAGACTT-3')-phosphate + L-tyrosine
-
 ?

Synonyms

Synonyms Comment Organism
TDP1
-
 Saccharomyces cerevisiae
tyrosyl-DNA phosphodiesterase I
-
 Saccharomyces cerevisiae

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.003
-
 (5'-GATCTAAAAGACTT-3')-phosphotyrosine mutant enzyme H182A, at pH 7.5 and 30°C Saccharomyces cerevisiae
0.007
-
 (5'-GATCTAAAAGACTT-3')-phosphotyrosine mutant enzyme H432R, at pH 7.5 and 30°C Saccharomyces cerevisiae
0.012
-
 (5'-GATCTAAAAGACTT-3')-phosphotyrosine mutant enzyme H432N, at pH 7.5 and 30°C Saccharomyces cerevisiae
0.28
-
 (5'-GATCTAAAAGACTT-3')-phosphotyrosine wild type enzyme, at pH 7.5 and 30°C Saccharomyces cerevisiae

General Information

General Information Comment Organism
physiological function the enzyme catalyzes the repair of 3'-DNA adducts, such as the 3'-phosphotyrosyl linkage of DNA topoisomerase I to DNA Saccharomyces cerevisiae

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.9
-
 (5'-GATCTAAAAGACTT-3')-phosphotyrosine mutant enzyme H182A, at pH 7.5 and 30°C Saccharomyces cerevisiae
1.5
-
 (5'-GATCTAAAAGACTT-3')-phosphotyrosine mutant enzyme H432R, at pH 7.5 and 30°C Saccharomyces cerevisiae
4.7
-
 (5'-GATCTAAAAGACTT-3')-phosphotyrosine mutant enzyme H432N, at pH 7.5 and 30°C Saccharomyces cerevisiae
250
-
 (5'-GATCTAAAAGACTT-3')-phosphotyrosine wild type enzyme, at pH 7.5 and 30°C Saccharomyces cerevisiae