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Literature summary for 3.6.4.B7 extracted from

  • Spies, M.; Kil, Y.; Masui, R.; Kato, R.; Kujo, C.; Ohshima, T.; Kuramitsu, S.; Lanzov, V.
    The RadA protein from a hyperthermophilic archaeon Pyrobaculum islandicum is a DNA-dependent ATPase that exhibits two disparate catalytic modes, with a transition temperature at 75°C (2000), Eur. J. Biochem., 267, 1125-1137.
    View publication on PubMed
Search for more literature for 3.6.4.B7

Activating Compound

Activating Compound Comment Organism Structure
ssDNA at 75°C, all DNAs tested stimulated ATPase activity of the RadA. The ssDNA-dependent ATPase activity of displays a temperature-dependent capacity to exist in two different catalytic modes, with 75°C being the critical threshold temperature Pyrobaculum islandicum

Cloned(Commentary)

Cloned (Comment) Organism
-
 Pyrobaculum islandicum

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
49
-
 ATP pH 7.7, 80°C, 0.02 mM poly(dT) Pyrobaculum islandicum
53
-
 ATP pH 7.7, 82°C, 0.02 mM poly(dT) Pyrobaculum islandicum
54
-
 ATP pH 7.7, 75°C, 0.02 mM poly(dC) Pyrobaculum islandicum
54
-
 ATP pH 7.7, 85°C, 0.02 mM poly(dT) Pyrobaculum islandicum
54
-
 ATP pH 7.7, °C, 0.02 mM M13mp18ssDNA Pyrobaculum islandicum
57
-
 ATP pH 7.7, °C, 0.02 mM M13mp19dsDNA Pyrobaculum islandicum
65
-
 ATP pH 7.7, 75°C, 0.02 mM poly(dT) Pyrobaculum islandicum
96
-
 ATP pH 7.7, 70°C, 0.02 mM poly(dT) Pyrobaculum islandicum
121
-
 ATP pH 7.7, 90°C, 0.02 mM poly(dT) Pyrobaculum islandicum
122
-
 ATP pH 7.7, 75°C, 0.02 mM poly(dA) Pyrobaculum islandicum
123
-
 ATP pH 7.7, 65°C, 0.02 mM poly(dT) Pyrobaculum islandicum
409
-
 ATP pH 7.7, 55°C, 0.02 mM poly(dT) Pyrobaculum islandicum
484
-
 ATP pH 7.7, 75°C, 0.02 mM DNA Pyrobaculum islandicum
531
-
 ATP pH 7.7, 37°C, 0.02 mM poly(dT) Pyrobaculum islandicum

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ the enzyme requires the presence of bivalent cations, such as Mg2+ and Mn2+ Pyrobaculum islandicum
Mn2+ the enzyme requires the presence of bivalent cations, such as Mg2+ and Mn2+ Pyrobaculum islandicum

Organic Solvent Stability

Organic Solvent Comment Organism
guanidine-HCl the enzyme is stable up to 4.5 M guanidine hydrochloride Pyrobaculum islandicum

Organism

Organism UniProt Comment Textmining
Pyrobaculum islandicum Q9UWR5
-
-
Pyrobaculum islandicum DSM 4184 Q9UWR5
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Pyrobaculum islandicum

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + H2O the enzyme binds ssDNA, hydrolyzes ATP in a DNA-dependent manner and to catalyzes DNA strand exchange.It shows the ability to bind ssDNA and catalyze DNA strand exchange between ssDNA and homologous linear dsDNA. The ssDNA-dependent ATPase activity displays a temperature-dependent capacity to exist in two different catalytic modes, with 75°C being the critical threshold temperature Pyrobaculum islandicum ADP + phosphate
-
 ?
ATP + H2O the enzyme binds ssDNA, hydrolyzes ATP in a DNA-dependent manner and to catalyzes DNA strand exchange.It shows the ability to bind ssDNA and catalyze DNA strand exchange between ssDNA and homologous linear dsDNA. The ssDNA-dependent ATPase activity displays a temperature-dependent capacity to exist in two different catalytic modes, with 75°C being the critical threshold temperature Pyrobaculum islandicum DSM 4184 ADP + phosphate
-
 ?

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
85
-
 the rate of the ATP hydrolysis increases with temperature up to 85°C, the maximum temperature at which the protein maintains its secondary structure Pyrobaculum islandicum

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
75
-
 the enzyme exhibits a biphasic Arrhenius plot of ATP hydrolysis with two characteristic energies of activation with a break point at 75°C. The activation energy below 75°C is higher than that above the break point. The cooperativity of ATP hydrolysis and ssDNA-binding ability of the protein above 75°C are greater than those at lower temperatures Pyrobaculum islandicum

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
25 87 the enzyme maintains its secondary structure and activities in vitro at high temperatures, up to 87°C. It also shows high stability of 18.3 kcal/mol at 25°C and neutral pH Pyrobaculum islandicum

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.013
-
 ATP pH 7.7, °C, 0.02 mM M13mp19dsDNA Pyrobaculum islandicum
0.015
-
 ATP pH 7.7, 37°C, 0.02 mM poly(dT) Pyrobaculum islandicum
0.018
-
 ATP pH 7.7, 75°C, 0.02 mM DNA Pyrobaculum islandicum
0.025
-
 ATP pH 7.7, 55°C, 0.02 mM poly(dT) Pyrobaculum islandicum
0.028
-
 ATP pH 7.7, 90°C, 0.02 mM poly(dT) Pyrobaculum islandicum
0.04
-
 ATP pH 7.7, 75°C, 0.02 mM poly(dA) Pyrobaculum islandicum
0.042
-
 ATP pH 7.7, 65°C, 0.02 mM poly(dT) Pyrobaculum islandicum
0.048
-
 ATP pH 7.7, 70°C, 0.02 mM poly(dT) Pyrobaculum islandicum
0.052
-
 ATP pH 7.7, °C, 0.02 mM M13mp18ssDNA Pyrobaculum islandicum
0.059
-
 ATP pH 7.7, 75°C, 0.02 mM poly(dT) Pyrobaculum islandicum
0.064
-
 ATP pH 7.7, 80°C, 0.02 mM poly(dT) Pyrobaculum islandicum
0.065
-
 ATP pH 7.7, 82°C, 0.02 mM poly(dT) Pyrobaculum islandicum
0.067
-
 ATP pH 7.7, 75°C, 0.02 mM poly(dC) Pyrobaculum islandicum
0.068
-
 ATP pH 7.7, 85°C, 0.02 mM poly(dT) Pyrobaculum islandicum