Activating Compound | Comment | Organism | Structure |
---|---|---|---|
Hexamine cobalt chloride | maximal stimulation at 0.001-0.0015 mM, blunt end ligation is stimulated 50fold, but cohesive end ligation only 5fold | Tequatrovirus T4 | |
Polyethylene glycol | polyethylene glycol 8000, maximal stimulation at 15% | Tequatrovirus T4 | |
Reducing agent | reducing agents, e.g. 2-mercaptoethanol or dithiothreitol required | Mammalia | |
Reducing agent | reducing agents, e.g. 2-mercaptoethanol or dithiothreitol required | Tequatrovirus T4 | |
Reducing agent | reducing agents, e.g. 2-mercaptoethanol or dithiothreitol required | Escherichia phage T7 | |
T4RNA ligase | stimulates T4 DNA ligase activity | Tequatrovirus T4 |
General Stability | Organism |
---|---|
a 10 U/ml dilution loses 40% of its activity in 3 months | Tequatrovirus T4 |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Cs+ | 0.2 M | Tequatrovirus T4 | |
dATP | - |
Escherichia phage T7 | |
dATP | - |
Mammalia | |
dATP | - |
Tequatrovirus T4 | |
K+ | 0.2 M | Tequatrovirus T4 | |
Li+ | 0.2 M | Tequatrovirus T4 | |
Na+ | 0.2 M | Tequatrovirus T4 | |
NH4+ | 0.2 M | Tequatrovirus T4 | |
phosphate | inhibits blunt end ligation | Tequatrovirus T4 | |
spermidine | - |
Tequatrovirus T4 | |
spermine | - |
Tequatrovirus T4 |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | - |
Mammalia | |
Mg2+ | optimal concentration: 10 mM | Tequatrovirus T4 | |
Mn2+ | 25% as effective as Mg2+ in activation | Tequatrovirus T4 |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
63000 | - |
1 * 63000, PAGE under denaturing and reducing conditions | Tequatrovirus T4 |
68000 | - |
gel filtration | Tequatrovirus T4 |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia phage T7 | - |
- |
- |
Mammalia | - |
- |
- |
Tequatrovirus T4 | - |
- |
- |
Storage Stability | Organism |
---|---|
-20°C, high concentrations of enzyme are very stable | Tequatrovirus T4 |
-20°C, storage below -20°C is detrimental | Tequatrovirus T4 |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + (deoxyribonucleotide)n + (deoxyribonucleotide)m | - |
Mammalia | AMP + diphosphate + (deoxyribonucleotide)n+m | - |
? | |
ATP + (deoxyribonucleotide)n + (deoxyribonucleotide)m | - |
Escherichia phage T7 | AMP + diphosphate + (deoxyribonucleotide)n+m | - |
? | |
ATP + (deoxyribonucleotide)n + (deoxyribonucleotide)m | low DNA concentrations favor intramolecular reaction, i.e. recircularization, and higher concentrations favor intermolecular reaction, i.e. oligomerization and formation of recombinant molecules | Tequatrovirus T4 | AMP + diphosphate + (deoxyribonucleotide)n+m | - |
? | |
ATP + (deoxyribonucleotide)n + (deoxyribonucleotide)m | some activity in joining RNA molecules annealed to DNA and even RNA:RNA molecules | Tequatrovirus T4 | AMP + diphosphate + (deoxyribonucleotide)n+m | - |
? | |
ATP + (deoxyribonucleotide)n + (deoxyribonucleotide)m | catalyzes blunt end joining of DNA | Tequatrovirus T4 | AMP + diphosphate + (deoxyribonucleotide)n+m | - |
? | |
ATP + (deoxyribonucleotide)n + (deoxyribonucleotide)m | reverse reaction is catalyzed, the enzyme behaves as an AMP-dependent endonuclease, yielding nicked DNA | Tequatrovirus T4 | AMP + diphosphate + (deoxyribonucleotide)n+m | - |
? | |
ATP + (deoxyribonucleotide)n + (deoxyribonucleotide)m | ligation of flush-ended DNAs | Tequatrovirus T4 | AMP + diphosphate + (deoxyribonucleotide)n+m | - |
? | |
additional information | - |
Mammalia | ? | - |
? | |
additional information | ATP-diphosphate exchange reaction | Tequatrovirus T4 | ? | - |
? | |
additional information | ATP-diphosphate exchange reaction | Escherichia phage T7 | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
monomer | 1 * 63000, PAGE under denaturing and reducing conditions | Tequatrovirus T4 |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
4 | - |
ligation of cohesive ends | Tequatrovirus T4 |
37 | - |
sealing nicks | Tequatrovirus T4 |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6.5 | - |
ATP-diphosphate exchange reaction | Tequatrovirus T4 |
7.2 | 7.8 | joining of nicks in Tris-HCl buffer | Tequatrovirus T4 |
7.5 | 8 | joining of DNA | Tequatrovirus T4 |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
6.9 | 8.3 | 6.9: 40% of maximal activity, 8.3: 65% of maximal activity, joing of DNA | Tequatrovirus T4 |