Cloned (Comment) | Organism |
---|---|
recombinant expression of His-tagged human APTX in Escherichia coli strain Rosetta 2 (DE3) cells | Homo sapiens |
Crystallization (Comment) | Organism |
---|---|
purified wild-type enzyme in complex with RNA/DNA, hAPTX-nicked-RNA-DNA-AMP-Zn2+ complex is grown by mixing 300 nl of 10 mg/ml hAptx (165-342) protein and 1 mM AMP, 1.5:1 DNA:protein molar ratio, in 150 mM NaCl, 20 mM Tris-HCl, pH 7.5, and 0.1% 2-mercaptoethanol, with an equal volume of precipitant solution containing 100 mM MES, pH 6.5, 16% w/v PEG 3350 at 4°C, methods for mutant product complexes hAPTX-R199H/RNA-DNA/AMP-Zn2+, hAPTXH201Q/RNA-DNA/AMP-Zn2+, hAPTX-S242N/RNA-DNA/AMPZn2+, hAPTX-L248M/DNA/AMP-Zn2+, and hAPTX-V263G/RNA-DNA/AMP-Zn2+, differ, overview. X-ray diffraction structure determination and analysis, molecular replacement using PDB ID 4NDG as a search model | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
A198V | site-directed mutagenesis, a mutation causing the neurodegenerative disorder ataxia with oculomotor ataxia 1 (AOA1) | Homo sapiens |
A198V/P206L | site-directed mutagenesis, a mutation causing the neurodegenerative disorder ataxia with oculomotor ataxia 1 (AOA1) | Homo sapiens |
D185E | site-directed mutagenesis, a mutation causing the neurodegenerative disorder ataxia with oculomotor ataxia 1 (AOA1) | Homo sapiens |
D267G | site-directed mutagenesis, a mutation causing the neurodegenerative disorder ataxia with oculomotor ataxia 1 (AOA1) | Homo sapiens |
D267G/W279X | site-directed mutagenesis, a mutation causing the neurodegenerative disorder ataxia with oculomotor ataxia 1 (AOA1) | Homo sapiens |
G231E | site-directed mutagenesis, a mutation causing the neurodegenerative disorder ataxia with oculomotor ataxia 1 (AOA1) | Homo sapiens |
G231E/689insT | site-directed mutagenesis, a mutation causing the neurodegenerative disorder ataxia with oculomotor ataxia 1 (AOA1) | Homo sapiens |
H201Q | site-directed mutagenesis, a mutation causing the neurodegenerative disorder ataxia with oculomotor ataxia 1 (AOA1) | Homo sapiens |
H201R | site-directed mutagenesis, a mutation causing the neurodegenerative disorder ataxia with oculomotor ataxia 1 (AOA1) | Homo sapiens |
K197Q | site-directed mutagenesis, a mutation causing the neurodegenerative disorder ataxia with oculomotor ataxia 1 (AOA1) | Homo sapiens |
K197Q/W279X | site-directed mutagenesis, a mutation causing the neurodegenerative disorder ataxia with oculomotor ataxia 1 (AOA1) | Homo sapiens |
L223P | site-directed mutagenesis, a mutation causing the neurodegenerative disorder ataxia with oculomotor ataxia 1 (AOA1) | Homo sapiens |
L248M | site-directed mutagenesis, a mutation causing the neurodegenerative disorder ataxia with oculomotor ataxia 1 (AOA1) | Homo sapiens |
R199H | site-directed mutagenesis, a mutation causing the neurodegenerative disorder ataxia with oculomotor ataxia 1 (AOA1) | Homo sapiens |
R247X | site-directed mutagenesis, not involved in AOA1 disease | Homo sapiens |
R306X | site-directed mutagenesis, a mutation causing the neurodegenerative disorder ataxia with oculomotor ataxia 1 (AOA1) | Homo sapiens |
R306X/W279X | site-directed mutagenesis, a mutation causing the neurodegenerative disorder ataxia with oculomotor ataxia 1 (AOA1) | Homo sapiens |
S242N | site-directed mutagenesis, a mutation causing the neurodegenerative disorder ataxia with oculomotor ataxia 1 (AOA1) | Homo sapiens |
V263G | site-directed mutagenesis, a mutation causing the neurodegenerative disorder ataxia with oculomotor ataxia 1 (AOA1) | Homo sapiens |
V263G/P206L | site-directed mutagenesis, a mutation causing the neurodegenerative disorder ataxia with oculomotor ataxia 1 (AOA1) | Homo sapiens |
W279R | site-directed mutagenesis, a mutation causing the neurodegenerative disorder ataxia with oculomotor ataxia 1 (AOA1) | Homo sapiens |
W279R/IVS5 | site-directed mutagenesis, a mutation causing the neurodegenerative disorder ataxia with oculomotor ataxia 1 (AOA1) | Homo sapiens |
W279X | site-directed mutagenesis, a mutation causing the neurodegenerative disorder ataxia with oculomotor ataxia 1 (AOA1) | Homo sapiens |
W279X/I159fs | site-directed mutagenesis, a mutation causing the neurodegenerative disorder ataxia with oculomotor ataxia 1 (AOA1) | Homo sapiens |
W279X/Q181X | site-directed mutagenesis, a mutation causing the neurodegenerative disorder ataxia with oculomotor ataxia 1 (AOA1) | Homo sapiens |
W279X/R306X | site-directed mutagenesis, a mutation causing the neurodegenerative disorder ataxia with oculomotor ataxia 1 (AOA1) | Homo sapiens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0000171 | - |
adenosine-5'-diphospho-5'-[DNA] | nicked DNA substrate, pH 7.5, 25-37°C, recombinant His-tagged wild-type enzyme | Homo sapiens | |
0.0000371 | - |
adenosine-5'-diphospho-5'-[DNA] | blunt DNA substrate, pH 7.5, 25-37°C, recombinant His-tagged wild-type enzyme | Homo sapiens |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Zn2+ | required | Homo sapiens |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
adenosine-5'-diphospho-5'-[DNA] + H2O | Homo sapiens | - |
AMP + phospho-5'-[DNA] | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | Q7Z2E3 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged human APTX from Escherichia coli strain Rosetta 2 (DE3) cells by nickel affinity chromatography | Homo sapiens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
adenosine-5'-diphospho-5'-[DNA] + H2O | - |
Homo sapiens | AMP + phospho-5'-[DNA] | - |
? | |
adenosine-5'-diphospho-5'-[DNA] + H2O | the enzyme can use nicked and blunt substrates | Homo sapiens | AMP + phospho-5'-[DNA] | - |
? | |
additional information | substrate binding structure, overview | Homo sapiens | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
APTX | - |
Homo sapiens |
RNA-DNA deadenylase | - |
Homo sapiens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | 37 | assay at | Homo sapiens |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.38 | - |
adenosine-5'-diphospho-5'-[DNA] | nicked DNA substrate, pH 7.5, 25-37°C, recombinant His-tagged wild-type enzyme | Homo sapiens | |
0.51 | - |
adenosine-5'-diphospho-5'-[DNA] | blunt DNA substrate, pH 7.5, 25-37°C, recombinant His-tagged wild-type enzyme | Homo sapiens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Homo sapiens |
General Information | Comment | Organism |
---|---|---|
malfunction | APTX human mutations cause the neurodegenerative disorder ataxia with oculomotor ataxia 1 (AOA1). AOA1 mutagenic effects on APTX solubility, stability, and catalytic activity, and molecular basis for APTX inactivation in AOA1, APTX mutations variably impact protein folding and activity, overview | Homo sapiens |
physiological function | the APTX RNA-DNA deadenylase protects genome integrity and corrects abortive DNA ligation arising during ribonucleotide excision repair and base excision DNA repair, APTX nicked DNA sensing and pleiotropic inactivation in neurodegenerative disease, mechanism | Homo sapiens |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
13747 | - |
adenosine-5'-diphospho-5'-[DNA] | blunt DNA substrate, pH 7.5, 25-37°C, recombinant His-tagged wild-type enzyme | Homo sapiens | |
22222 | - |
adenosine-5'-diphospho-5'-[DNA] | nicked DNA substrate, pH 7.5, 25-37°C, recombinant His-tagged wild-type enzyme | Homo sapiens |