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Literature summary for 6.5.1.1 extracted from

  • Maunders, M.J.
    DNA and RNA ligases (EC 6.5.1.1, EC 6.5.1.2, and EC 6.5.1.3) (1993), Methods Mol. Biol., 16, 213-230.
    View publication on PubMed
Search for more literature for 6.5.1.1

Activating Compound

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

General Stability Organism
a 10 U/ml dilution loses 40% of its activity in 3 months Tequatrovirus T4

Inhibitors

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

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 [Da] Molecular Weight Maximum [Da] Comment Organism
63000
-
 1 * 63000, PAGE under denaturing and reducing conditions Tequatrovirus T4
68000
-
 gel filtration Tequatrovirus T4

Organism

Organism UniProt Comment Textmining
Escherichia phage T7
-
-
-
Mammalia
-
-
-
Tequatrovirus T4
-
-
-

Storage Stability

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 and Products (Substrate)

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

Subunits Comment Organism
monomer 1 * 63000, PAGE under denaturing and reducing conditions Tequatrovirus T4

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
4
-
 ligation of cohesive ends Tequatrovirus T4
37
-
 sealing nicks Tequatrovirus T4

pH Optimum

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 Range

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