Application | Comment | Organism |
---|---|---|
additional information | opposite-base specificity of Nei is primarily governed by residues in the Q69-Y71 loop, both Q69-Y71 loop and the zinc finger contribute significantly to the substrate specificity of Nei | Escherichia coli |
Cloned (Comment) | Organism |
---|---|
into vector pET-24b, recombinant plasmids maintained in Escherichia coli XL1-Blue X cells, transfected into Escherichia L21(DE3) | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
deltaQLY69-71 | deletion of the entire loop, interferes with eversion of the damaged base from the helix, deficient in processing damaged DNA | Escherichia coli |
Q261A | perturbs the conserved zinc finger, deficient in processing damaged DNA, can be reductively cross-linked to damaged base-containing DNA, deficient in regenerating free enzyme from the Nei-DNA covalent complex formed during the reaction | Escherichia coli |
QLY69-71AAA | all amino acids in the QLY loop substituted with alanines, interferes with eversion of the damaged base from the helix, deficient in processing damaged DNA | Escherichia coli |
R171A | perturbs the conserved zinc finger, deficient in processing damaged DNA | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.000000083 | - |
DNA containing apurinic/apyrimidinic sites | wild-type, Nei placed opposite A | Escherichia coli | |
0.00000011 | - |
DNA containing apurinic/apyrimidinic sites | wild-type, Nei placed opposite G | Escherichia coli | |
0.0000023 | - |
5'-GACAAGCGCAG-(5R,6S)-2'-deoxy-5,6-dihydroxyuridine-CAGCCGAACAC-3' | wild-type, Nei placed opposite G | Escherichia coli | |
0.0000023 | - |
5'-GACAAGCGCAG-(5S,6R)-2'-deoxy-5,6-dihydroxyuridine-CAGCCGAACAC-3' | wild-type, Nei placed opposite G | Escherichia coli | |
0.0000032 | - |
5'-GACAAGCGCAG-(5R,6S)-2'-deoxy-5,6-dihydroxyuridine-CAGCCGAACAC-3' | wild-type, Nei placed opposite A | Escherichia coli | |
0.0000035 | - |
5'-CTCTCCCTTC-5,6-dihydrouracil-CTCCTTTCCTCT-3' | wild-type, Nei placed opposite T | Escherichia coli | |
0.0000044 | - |
5'-GACAAGCGCAG-(5R,6S)-2'-deoxy-5,6-dihydroxyuridine-CAGCCGAACAC-3' | wild-type, Nei placed opposite C | Escherichia coli | |
0.0000044 | - |
5'-GACAAGCGCAG-(5R,6S)-2'-deoxy-5,6-dihydroxyuridine-CAGCCGAACAC-3' | wild-type, Nei placed opposite T | Escherichia coli | |
0.0000046 | - |
5'-GACAAGCGCAG-(5S,6R)-2'-deoxy-5,6-dihydroxyuridine-CAGCCGAACAC-3' | wild-type, Nei placed opposite T | Escherichia coli | |
0.0000047 | - |
5'-GACAAGCGCAG-(5S,6R)-2'-deoxy-5,6-dihydroxyuridine-CAGCCGAACAC-3' | wild-type, Nei placed opposite C | Escherichia coli | |
0.0000066 | - |
5'-CTCTCCCTTC-5,6-dihydrouracil-CTCCTTTCCTCT-3' | wild-type, Nei placed opposite C | Escherichia coli | |
0.0000085 | - |
5'-CTCTCCCTTC-5,6-dihydrouracil-CTCCTTTCCTCT-3' | wild-type, Nei placed opposite G | Escherichia coli | |
0.000016 | - |
5'-GACAAGCGCAG-(5S,6R)-2'-deoxy-5,6-dihydroxyuridine-CAGCCGAACAC-3' | wild-type, Nei placed opposite A | Escherichia coli | |
0.000022 | - |
DNA containing apurinic/apyrimidinic sites | mutant QLY/AAA, Nei placed opposite G | Escherichia coli | |
0.000028 | - |
DNA containing apurinic/apyrimidinic sites | mutant QLY/AAA, Nei placed opposite A | Escherichia coli | |
0.000038 | - |
5'-CTCTCCCTTC-5,6-dihydrouracil-CTCCTTTCCTCT-3' | mutant QLY/AAA, Nei placed opposite A | Escherichia coli | |
0.000042 | - |
5'-CTCTCCCTTC-5,6-dihydrouracil-CTCCTTTCCTCT-3' | mutant QLY/AAA, Nei placed opposite G | Escherichia coli | |
0.000049 | - |
5'-GACAAGCGCAG-(5R,6S)-2'-deoxy-5,6-dihydroxyuridine-CAGCCGAACAC-3' | mutant QLY/AAA, Nei placed opposite G | Escherichia coli | |
0.000054 | - |
5'-CTCTCCCTTC-5,6-dihydrouracil-CTCCTTTCCTCT-3' | mutant deltaQLY, Nei placed opposite G | Escherichia coli | |
0.000066 | - |
5'-GACAAGCGCAG-(5R,6S)-2'-deoxy-5,6-dihydroxyuridine-CAGCCGAACAC-3' | mutant deltaQLY, Nei placed opposite A | Escherichia coli | |
0.000077 | - |
5'-CTCTCCCTTC-5,6-dihydrouracil-CTCCTTTCCTCT-3' | mutant Q261A, Nei placed opposite G | Escherichia coli | |
0.00014 | - |
5'-CTCTCCCTTC-5,6-dihydrouracil-CTCCTTTCCTCT-3' | mutant QLY/AAA, Nei placed opposite C | Escherichia coli | |
0.00016 | - |
5'-CTCTCCCTTC-5,6-dihydrouracil-CTCCTTTCCTCT-3' | mutant QLY/AAA, Nei placed opposite T | Escherichia coli | |
0.00016 | - |
5'-CTCTCCCTTC-5,6-dihydrouracil-CTCCTTTCCTCT-3' | wild-type, Nei placed opposite A | Escherichia coli | |
0.00019 | - |
5'-GACAAGCGCAG-(5R,6S)-2'-deoxy-5,6-dihydroxyuridine-CAGCCGAACAC-3' | mutant QLY/AAA, Nei placed opposite T | Escherichia coli | |
0.00021 | - |
5'-GACAAGCGCAG-(5S,6R)-2'-deoxy-5,6-dihydroxyuridine-CAGCCGAACAC-3' | mutant Q261A, Nei placed opposite G | Escherichia coli | |
0.00024 | - |
5'-GACAAGCGCAG-(5R,6S)-2'-deoxy-5,6-dihydroxyuridine-CAGCCGAACAC-3' | mutant Q261A, Nei placed opposite G | Escherichia coli | |
0.00026 | - |
5'-CTCTCCCTTC-5,6-dihydrouracil-CTCCTTTCCTCT-3' | mutant Q261A, Nei placed opposite A | Escherichia coli | |
0.00027 | - |
5'-CTCTCCCTTC-5,6-dihydrouracil-CTCCTTTCCTCT-3' | mutant Q261A, Nei placed opposite C | Escherichia coli | |
0.00058 | - |
5'-CTCTCCCTTC-8-oxo-7,8-dihydroguanine-CTCCTTTCCTCT-3' | wild-type, Nei placed opposite G | Escherichia coli | |
0.00061 | - |
5'-CTCTCCCTTC-8-oxo-7,8-dihydroguanine-CTCCTTTCCTCT-3' | wild-type, Nei placed opposite A | Escherichia coli | |
0.00068 | - |
5'-CTCTCCCTTC-8-oxo-7,8-dihydroguanine-CTCCTTTCCTCT-3' | wild-type, Nei placed opposite C | Escherichia coli | |
0.0013 | - |
5'-CTCTCCCTTC-8-oxo-7,8-dihydroguanine-CTCCTTTCCTCT-3' | wild-type, Nei placed opposite T | Escherichia coli | |
0.0014 | - |
5'-CTCTCCCTTC-5,6-dihydrouracil-CTCCTTTCCTCT-3' | mutant Q261A, Nei placed opposite T | Escherichia coli |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Zn2+ | - |
Escherichia coli |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | P50465 | - |
- |
Purification (Comment) | Organism |
---|---|
- |
Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
5'-CTCTCCCTTC-5,6-dihydrouracil-CTCCTTTCCTCT-3' | - |
Escherichia coli | ? | - |
? | |
5'-CTCTCCCTTC-8-oxo-7,8-dihydroguanine-CTCCTTTCCTCT-3' | - |
Escherichia coli | ? | - |
? | |
5'-GACAAGCGCAG-(5R,6S)-2'-deoxy-5,6-dihydroxyuridine-CAGCCGAACAC-3' | - |
Escherichia coli | ? | - |
? | |
5'-GACAAGCGCAG-(5S,6R)-2'-deoxy-5,6-dihydroxyuridine-CAGCCGAACAC-3' | - |
Escherichia coli | ? | - |
? | |
DNA containing apurinic/apyrimidinic sites | - |
Escherichia coli | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
endonuclease VIII | - |
Escherichia coli |
Nei | - |
Escherichia coli |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0000048 | - |
5'-CTCTCCCTTC-5,6-dihydrouracil-CTCCTTTCCTCT-3' | mutant deltaQLY, Nei placed opposite G | Escherichia coli | |
0.000013 | - |
5'-CTCTCCCTTC-5,6-dihydrouracil-CTCCTTTCCTCT-3' | mutant Q261A, Nei placed opposite C | Escherichia coli | |
0.000015 | - |
5'-CTCTCCCTTC-5,6-dihydrouracil-CTCCTTTCCTCT-3' | mutant Q261A, Nei placed opposite G | Escherichia coli | |
0.000023 | - |
5'-CTCTCCCTTC-5,6-dihydrouracil-CTCCTTTCCTCT-3' | mutant Q261A, Nei placed opposite A | Escherichia coli | |
0.000065 | - |
5'-CTCTCCCTTC-5,6-dihydrouracil-CTCCTTTCCTCT-3' | mutant QLY/AAA, Nei placed opposite G | Escherichia coli | |
0.000067 | - |
5'-CTCTCCCTTC-5,6-dihydrouracil-CTCCTTTCCTCT-3' | mutant Q261A, Nei placed opposite T | Escherichia coli | |
0.00007167 | - |
5'-CTCTCCCTTC-5,6-dihydrouracil-CTCCTTTCCTCT-3' | mutant QLY/AAA, Nei placed opposite A | Escherichia coli | |
0.000583 | - |
5'-CTCTCCCTTC-5,6-dihydrouracil-CTCCTTTCCTCT-3' | wild-type, Nei placed opposite A | Escherichia coli | |
0.0015 | - |
DNA containing apurinic/apyrimidinic sites | mutant QLY/AAA, Nei placed opposite A | Escherichia coli | |
0.00183 | - |
DNA containing apurinic/apyrimidinic sites | mutant QLY/AAA, Nei placed opposite G | Escherichia coli | |
0.0025 | - |
5'-GACAAGCGCAG-(5R,6S)-2'-deoxy-5,6-dihydroxyuridine-CAGCCGAACAC-3' | mutant deltaQLY, Nei placed opposite A | Escherichia coli | |
0.003167 | - |
5'-CTCTCCCTTC-5,6-dihydrouracil-CTCCTTTCCTCT-3' | wild-type, Nei placed opposite T | Escherichia coli | |
0.00567 | - |
5'-GACAAGCGCAG-(5S,6R)-2'-deoxy-5,6-dihydroxyuridine-CAGCCGAACAC-3' | mutant Q261A, Nei placed opposite G | Escherichia coli | |
0.00583 | - |
5'-GACAAGCGCAG-(5R,6S)-2'-deoxy-5,6-dihydroxyuridine-CAGCCGAACAC-3' | mutant Q261A, Nei placed opposite G | Escherichia coli | |
0.00783 | - |
5'-CTCTCCCTTC-8-oxo-7,8-dihydroguanine-CTCCTTTCCTCT-3' | wild-type, Nei placed opposite A | Escherichia coli | |
0.008167 | - |
5'-CTCTCCCTTC-5,6-dihydrouracil-CTCCTTTCCTCT-3' | wild-type, Nei placed opposite C | Escherichia coli | |
0.00883 | - |
DNA containing apurinic/apyrimidinic sites | wild-type, Nei placed opposite G | Escherichia coli | |
0.01067 | - |
DNA containing apurinic/apyrimidinic sites | wild-type, Nei placed opposite A | Escherichia coli | |
0.0112 | - |
5'-GACAAGCGCAG-(5S,6R)-2'-deoxy-5,6-dihydroxyuridine-CAGCCGAACAC-3' | wild-type, Nei placed opposite T | Escherichia coli | |
0.0115 | - |
5'-GACAAGCGCAG-(5S,6R)-2'-deoxy-5,6-dihydroxyuridine-CAGCCGAACAC-3' | wild-type, Nei placed opposite C | Escherichia coli | |
0.01167 | - |
5'-CTCTCCCTTC-5,6-dihydrouracil-CTCCTTTCCTCT-3' | mutant QLY/AAA, Nei placed opposite T | Escherichia coli | |
0.013 | - |
5'-CTCTCCCTTC-5,6-dihydrouracil-CTCCTTTCCTCT-3' | mutant QLY/AAA, Nei placed opposite C | Escherichia coli | |
0.01317 | - |
5'-CTCTCCCTTC-8-oxo-7,8-dihydroguanine-CTCCTTTCCTCT-3' | wild-type, Nei placed opposite G | Escherichia coli | |
0.01583 | - |
5'-CTCTCCCTTC-8-oxo-7,8-dihydroguanine-CTCCTTTCCTCT-3' | wild-type, Nei placed opposite C | Escherichia coli | |
0.016 | - |
5'-GACAAGCGCAG-(5R,6S)-2'-deoxy-5,6-dihydroxyuridine-CAGCCGAACAC-3' | mutant QLY/AAA, Nei placed opposite G | Escherichia coli | |
0.02167 | - |
5'-GACAAGCGCAG-(5R,6S)-2'-deoxy-5,6-dihydroxyuridine-CAGCCGAACAC-3' | mutant QLY/AAA, Nei placed opposite T | Escherichia coli | |
0.023 | - |
5'-GACAAGCGCAG-(5S,6R)-2'-deoxy-5,6-dihydroxyuridine-CAGCCGAACAC-3' | wild-type, Nei placed opposite A | Escherichia coli | |
0.023 | - |
5'-CTCTCCCTTC-8-oxo-7,8-dihydroguanine-CTCCTTTCCTCT-3' | wild-type, Nei placed opposite T | Escherichia coli | |
0.03 | - |
5'-GACAAGCGCAG-(5R,6S)-2'-deoxy-5,6-dihydroxyuridine-CAGCCGAACAC-3' | wild-type, Nei placed opposite G | Escherichia coli | |
0.035 | - |
5'-GACAAGCGCAG-(5R,6S)-2'-deoxy-5,6-dihydroxyuridine-CAGCCGAACAC-3' | wild-type, Nei placed opposite A | Escherichia coli | |
0.04167 | - |
5'-GACAAGCGCAG-(5R,6S)-2'-deoxy-5,6-dihydroxyuridine-CAGCCGAACAC-3' | wild-type, Nei placed opposite C | Escherichia coli | |
0.0483 | - |
5'-GACAAGCGCAG-(5R,6S)-2'-deoxy-5,6-dihydroxyuridine-CAGCCGAACAC-3' | wild-type, Nei placed opposite T | Escherichia coli | |
0.0567 | - |
5'-GACAAGCGCAG-(5S,6R)-2'-deoxy-5,6-dihydroxyuridine-CAGCCGAACAC-3' | wild-type, Nei placed opposite G | Escherichia coli | |
0.07167 | - |
5'-CTCTCCCTTC-5,6-dihydrouracil-CTCCTTTCCTCT-3' | wild-type, Nei placed opposite G | Escherichia coli |