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Literature summary for 5.6.2.3 extracted from

  • Longley, M.J.; Humble, M.M.; Sharief, F.S.; Copeland, W.C.
    Disease variants of the human mitochondrial DNA helicase encoded by C10orf2 differentially alter protein stability, nucleotide hydrolysis, and helicase activity (2010), J. Biol. Chem., 285, 29690-29702.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 5.6.2.3

Cloned(Commentary)

Cloned (Comment) Organism
overexpressed in Escherichia coli as His-tagged fusion proteins Homo sapiens

Protein Variants

Protein Variants Comment Organism
A318T mutation associated with mitochondrial disease does not cause profound defects in DNA binding, DNA helicase function, or ATPase activity, kcat/Km (ATP) comparable to wild-type Homo sapiens
A359T mutation associated with mitochondrial disease does not cause profound defects in DNA binding, DNA helicase function, or ATPase activity, kcat/Km (ATP) decreased compared to wild-type, mutant shows a 3-4fold increase in Kd (dsDNA) value compared to wild-type, mutant is much more sensitive to heat inactivation than wild-type Homo sapiens
A475P mutation associated with mitochondrial disease does not cause profound defects in DNA binding, DNA helicase function, or ATPase activity, kcat/Km (ATP) decreased compared to wild-type, mutant shows lowest kcat (ATP) value Homo sapiens
F485L mutation associated with mitochondrial disease does not cause profound defects in DNA binding, DNA helicase function, or ATPase activity, kcat/Km (ATP) decreased compared to wild-type, mutant shows a 3-4fold increase in Kd (dsDNA) value compared to wild-type, mutant is much more sensitive to heat inactivation than wild-type Homo sapiens
I367T mutation associated with mitochondrial disease does not cause profound defects in DNA binding, DNA helicase function, or ATPase activity, kcat/Km (ATP) slightly decreased compared to wild-type, mutant shows a higher Km (ATP) and kcat value compared to wild-type Homo sapiens
K319E mutation associated with mitochondrial disease does not cause profound defects in DNA binding, DNA helicase function, or ATPase activity, kcat/Km (ATP) comparable to wild-type Homo sapiens
K319T mutation associated with mitochondrial disease does not cause profound defects in DNA binding, DNA helicase function, or ATPase activity, kcat/Km (ATP) decreased compared to wild-type Homo sapiens
L381P mutation associated with mitochondrial disease does not cause profound defects in DNA binding, DNA helicase function, or ATPase activity, kcat/Km (ATP) increased compared to wild-type, mutant shows a higher Km (ATP) and kcat value compared to wild-type Homo sapiens
P335L mutation associated with mitochondrial disease does not cause profound defects in DNA binding, DNA helicase function, or ATPase activity, kcat/Km (ATP) decreased compared to wild-type Homo sapiens
R303W mutation associated with mitochondrial disease does not cause profound defects in DNA binding, DNA helicase function, or ATPase activity, kcat/Km (ATP) comparable to wild-type, mutant is much more sensitive to heat inactivation than wild-type Homo sapiens
R334Q mutation associated with mitochondrial disease does not cause profound defects in DNA binding, DNA helicase function, or ATPase activity, kcat/Km (ATP) decreased compared to wild-type Homo sapiens
R354P mutation associated with mitochondrial disease does not cause profound defects in DNA binding, DNA helicase function, or ATPase activity, kcat/Km (ATP) decreased compared to wild-type Homo sapiens
R357P mutation associated with mitochondrial disease does not cause profound defects in DNA binding, DNA helicase function, or ATPase activity, kcat/Km (ATP) slightly decreased compared to wild-type Homo sapiens
R374Q mutation associated with mitochondrial disease does not cause profound defects in DNA binding, DNA helicase function, or ATPase activity, kcat/Km (ATP) decreased compared to wild-type, mutant shows a higher Km (ATP) value compared to wild-type, mutant shows a 3-4fold increase in Kd (dsDNA) and a 2fold increase in Kd (ssDNA) value compared to wild-type, mutant is much more sensitive to heat inactivation than wild-type Homo sapiens
S369P mutation associated with mitochondrial disease does not cause profound defects in DNA binding, DNA helicase function, or ATPase activity, kcat/Km (ATP) decreased compared to wild-type Homo sapiens
S369Y mutation associated with mitochondrial disease does not cause profound defects in DNA binding, DNA helicase function, or ATPase activity, kcat/Km (ATP) increased compared to wild-type, mutant shows a 3-4fold increase in Kd (dsDNA) value compared to wild-type, mutant is much more sensitive to heat inactivation than wild-type Homo sapiens
T457I mutation associated with mitochondrial disease does not cause profound defects in DNA binding, DNA helicase function, or ATPase activity, kcat/Km (ATP) decreased compared to wild-type Homo sapiens
W315L mutation associated with mitochondrial disease does not cause profound defects in DNA binding, DNA helicase function, or ATPase activity, kcat/Km (ATP) slightly decreased compared to wild-type, mutant is much more sensitive to heat inactivation than wild-type Homo sapiens
W474C mutation associated with mitochondrial disease does not cause profound defects in DNA binding, DNA helicase function, or ATPase activity, kcat/Km (ATP) decreased compared to wild-type Homo sapiens
Y508C mutation associated with mitochondrial disease does not cause profound defects in DNA binding, DNA helicase function, or ATPase activity, kcat/Km (ATP) decreased compared to wild-type, mutant shows a 3-4fold increase in Kd (dsDNA) value compared to wild-type Homo sapiens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.84
-
 ATP mutant A318T, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
1
-
 ATP mutant W315L, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
1.02
-
 ATP mutant K319T, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
1.03
-
 ATP mutant R354P, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
1.04
-
 ATP mutant R334Q, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
1.08
-
 ATP mutant T457I, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
1.1
-
 ATP mutant W474C, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
1.17
-
 ATP mutant S369Y pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
1.22
-
 ATP mutant A475P, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
1.22
-
 ATP wild-type wit His-tag, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
1.24
-
 ATP mutant R357P, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
1.3
-
 ATP mutant A359T, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
1.32
-
 ATP mutant P335L, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
1.43
-
 ATP wild-type without His-tag, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
1.44
-
 ATP mutant F485L, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
1.81
-
 ATP mutant R303W, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
1.82
-
 ATP mutant K319E, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
1.89
-
 ATP mutant S369P, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
1.96
-
 ATP mutant Y508C, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
2.23
-
 ATP mutant I367T, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
2.48
-
 ATP mutant R374Q , pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
4.55
-
 ATP mutant L381P, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens

Localization

Localization Comment Organism GeneOntology No. Textmining
mitochondrion
-
 Homo sapiens 5739
-

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
72000
-
 SDS-PAGE Homo sapiens

Organism

Organism UniProt Comment Textmining
Homo sapiens Q96RR1
-
-

Purification (Commentary)

Purification (Comment) Organism
using Ni-NTA chromatography Homo sapiens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + H2O
-
 Homo sapiens ADP + phosphate
-
 ?

Subunits

Subunits Comment Organism
hexamer
-
 Homo sapiens

Synonyms

Synonyms Comment Organism
C10orf2
-
 Homo sapiens

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Homo sapiens

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
42
-
 wild-type protein has a half-liife if 7.9 min at 42°C Homo sapiens

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.66
-
 ATP mutant A475P, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
0.7
-
 ATP mutant W474C, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
0.88
-
 ATP mutant T457I, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
0.98
-
 ATP mutant K319T, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
1.01
-
 ATP mutant R334Q, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
1.06
-
 ATP mutant A359T, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
1.08
-
 ATP mutant P335L, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
1.13
-
 ATP mutant W315L, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
1.15
-
 ATP mutant F485L, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
1.18
-
 ATP mutant R354P, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
1.19
-
 ATP mutant A318T, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
1.52
-
 ATP mutant R357P, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
1.9
-
 ATP wild-type wit His-tag, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
1.94
-
 ATP mutant S369Y pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
2.04
-
 ATP mutant Y508C, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
2.09
-
 ATP mutant S369P, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
2.17
-
 ATP mutant R374Q , pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
2.18
-
 ATP wild-type without His-tag, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
2.24
-
 ATP mutant K319E, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
2.29
-
 ATP mutant R303W, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
2.89
-
 ATP mutant I367T, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
8.31
-
 ATP mutant L381P, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
 assay at Homo sapiens

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.54
-
 ATP mutant A475P, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
0.64
-
 ATP mutant W474C, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
0.8
-
 ATP mutant F485L, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
0.81
-
 ATP mutant A359T, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
0.81
-
 ATP mutant T457I, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
0.82
-
 ATP mutant P335L, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
0.88
-
 ATP mutant R374Q , pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
0.96
-
 ATP mutant K319T, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
0.97
-
 ATP mutant R334Q, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
1.04
-
 ATP mutant Y508C, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
1.11
-
 ATP mutant S369P, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
1.13
-
 ATP mutant W315L, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
1.15
-
 ATP mutant R354P, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
1.23
-
 ATP mutant K319E, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
1.23
-
 ATP mutant R357P, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
1.27
-
 ATP mutant R303W, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
1.3
-
 ATP mutant I367T, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
1.42
-
 ATP mutant A318T, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
1.53
-
 ATP wild-type without His-tag, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
1.55
-
 ATP wild-type wit His-tag, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
1.67
-
 ATP mutant S369Y pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens
1.83
-
 ATP mutant L381P, pH 7.5, 37°C, in the presence of calf thymus DNA Homo sapiens