Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli, pQE-80L vector used for generation of engineered variants containing a single amino acid substitution, His-tagging | Thermotoga maritima |
Crystallization (Comment) | Organism |
---|---|
four-layer alpha-beta/beta-alpha sandwich fold determined | Thermotoga maritima |
Protein Variants | Comment | Organism |
---|---|---|
D190A | in vitro 3'-processing in the presence of 10mM Mg2+ or 0.2 mM Mn2+ | Thermotoga maritima |
D25A | in vitro 3'-processing in the presence of 10mM Mg2+ or 0.2 mM Mn2+ | Thermotoga maritima |
D52A | in vitro 3'-processing in the presence of 10mM Mg2+ or 0.2 mM Mn2+ | Thermotoga maritima |
F11P | in vitro 3'-processing in the presence of 10mM Mg2+ or 0.2 mM Mn2+ | Thermotoga maritima |
H134A | in vitro 3'-processing in the presence of 10mM Mg2+ or 0.2 mM Mn2+ | Thermotoga maritima |
H222A | in vitro 3'-processing in the presence of 10mM Mg2+ or 0.2 mM Mn2+ | Thermotoga maritima |
H244A | in vitro 3'-processing in the presence of 10mM Mg2+ or 0.2 mM Mn2+ | Thermotoga maritima |
H48A | in vitro 3'-processing in the presence of 10mM Mg2+ or 0.2 mM Mn2+ | Thermotoga maritima |
H50A | in vitro 3'-processing in the presence of 10mM Mg2+ or 0.2 mM Mn2+ | Thermotoga maritima |
H53A | in vitro 3'-processing in the presence of 10mM Mg2+ or 0.2 mM Mn2+ | Thermotoga maritima |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Co2+ | 0.2 mM, no rescue of lost tRNase Z activity observed | Thermotoga maritima | |
Cu2+ | 0.2 mM, no rescue of lost tRNase Z activity observed | Thermotoga maritima | |
Mg2+ | 10 mM, reaction kinetics indicated, cleavage by each variant in the presence of Mg2+ hardly detected, binding to pre-tRNA observed, essential for activity | Thermotoga maritima | |
Mn2+ | 0.2 mM, reaction kinetics indicated, essential for activity, Mn2+ ions restore a lost Mg2+ dependent activity with two exceptions of the Asp52Ala and His222Ala substitutions | Thermotoga maritima | |
Zn2+ | 0.2 mM, no rescue of lost tRNase Z activity observed | Thermotoga maritima |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Thermotoga maritima | - |
- |
- |
Purification (Comment) | Organism |
---|---|
gel filtration | Thermotoga maritima |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
reaction kinetics for in vitro 3'-processing of pre-tRNAArg(74CCA76) and pre-tRNAArg(74GUG76) in the presence of 10mM Mg2+ or 0.2 mM Mn2+ shown | Thermotoga maritima |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
pre-tRNAArg(74CCA76) + H2O | reaction kinetics for in vitro 3'-processing using engineered enzyme variants, in presence of 10 mM Mg2+ or of 0.2 mM Mn2+ | Thermotoga maritima | ? | - |
? | |
pre-tRNAArg(74GUG76) + H2O | cleavage of pre-tRNAArg(74GUG76) by engineered variants about 7 to more than 16fold less efficient than that of pre-tRNAArg(74CCA76) | Thermotoga maritima | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
tRNase Z | - |
Thermotoga maritima |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
60 | - |
assay at | Thermotoga maritima |