Application | Comment | Organism |
---|---|---|
analysis | comparative studies both bis(p-nitrophenyl)phosphate hydrolysis and pre-tRNA processing of tRNase Z variants | Arabidopsis thaliana |
Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli | Drosophila melanogaster |
expressed in Escherichia coli | Saccharomyces cerevisiae |
expressed in Escherichia coli, prior to electrophoretic mobility shift assay (EMSA) and cross-linking experiments | Arabidopsis thaliana |
Protein Variants | Comment | Organism |
---|---|---|
C25G | variants of AthTRZ1 wild-type enzyme generated by mutagenesis | Arabidopsis thaliana |
C40G | variants of AthTRZ1 wild-type enzyme generated by mutagenesis | Arabidopsis thaliana |
E208A | variants of AthTRZ1 wild-type enzyme generated by mutagenesis | Arabidopsis thaliana |
F51L | variants of AthTRZ1 wild-type enzyme generated by mutagenesis | Arabidopsis thaliana |
G62V | variants of AthTRZ1 wild-type enzyme generated by mutagenesis | Arabidopsis thaliana |
P178A | variants of AthTRZ1 wild-type enzyme generated by mutagenesis | Arabidopsis thaliana |
P64A | variants of AthTRZ1 wild-type enzyme generated by mutagenesis | Arabidopsis thaliana |
R252G | variants of AthTRZ1 wild-type enzyme generated by mutagenesis | Arabidopsis thaliana |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
bis(p-nitrophenyl)phosphate | pre-tRNA processing inhibited by bis(p-nitrophenyl)phosphate concentrations higher than 10 mM observed | Drosophila melanogaster | |
bis(p-nitrophenyl)phosphate | pre-tRNA processing inhibited by bis(p-nitrophenyl)phosphate concentrations higher than 10 mM observed | Saccharomyces cerevisiae | |
additional information | reaction performed with 20 microM bis(p-nitrophenyl)phosphate and 20 microM tRNA, bis(p-nitrophenyl)phosphate does not inhibit tRNA binding | Arabidopsis thaliana |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.2 | - |
bis(p-nitrophenyl)phosphate | AthTRZ1-P178A, mutant, bis(p-nitrophenyl)phosphate catalysis | Arabidopsis thaliana | |
1.8 | - |
bis(p-nitrophenyl)phosphate | AthTRZ1-R252G, mutant, bis(p-nitrophenyl)phosphate catalysis | Arabidopsis thaliana | |
3.5 | - |
bis(p-nitrophenyl)phosphate | AthTRZ1-E208A, mutant, bis(p-nitrophenyl)phosphate catalysis | Arabidopsis thaliana | |
6 | - |
bis(p-nitrophenyl)phosphate | AthTRZ1-C40G, mutant, bis(p-nitrophenyl)phosphate catalysis | Arabidopsis thaliana | |
6.5 | - |
bis(p-nitrophenyl)phosphate | AthTRZ1-F51L, mutant, bis(p-nitrophenyl)phosphate catalysis | Arabidopsis thaliana | |
8 | - |
bis(p-nitrophenyl)phosphate | AthTRZ1-G62V, mutant, bis(p-nitrophenyl)phosphate catalysis | Arabidopsis thaliana | |
8.5 | - |
bis(p-nitrophenyl)phosphate | AthTRZ1 wild-type, hydrolysis of phosphodiester bonds of bpNPP, KM value for AthTRZ1 two-fold increased relative to that of the tRNase Z of Escherichia coli | Arabidopsis thaliana | |
13.2 | - |
bis(p-nitrophenyl)phosphate | AthTRZ1-C25G, mutant, bis(p-nitrophenyl)phosphate catalysis | Arabidopsis thaliana | |
22.2 | - |
bis(p-nitrophenyl)phosphate | AthTRZ1-P64A, mutant, bis(p-nitrophenyl)phosphate catalysis | Arabidopsis thaliana |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | requird for in Vitro tRNA processing reaction | Arabidopsis thaliana | |
Mg2+ | 2 mM, also required for in vitro tRNA processing reactions | Arabidopsis thaliana | |
Mn2+ | 0.2 mM, addition of Mn2+ ions triples activity, also required for in Vitro tRNA processing reactions | Arabidopsis thaliana | |
additional information | in contrast to bis(p-nitrophenyl)phosphate hydrolysis, pre-tRNA processing requires additional metal ions, Mn2+ or Mg2+, as Zn2+ ions alone are insufficient, metal dependence of the in vitro processing reaction analyzed, bis(p-nitrophenyl)phosphate activity of the chelator-treated AthTRZ1 without (TRZ-E) and with additional metal ions tested, chelator-treated AthTRZ1 without additional metal ions reveal same activity as the nonchelator-treated AthTRZ1 | Arabidopsis thaliana | |
Zn2+ | 0.2 mM, addition of Zn2+ ions to the chelator-treated enzyme doubles activity, high affinity to Zn2+ even upon incubation with metal chelators, 0.76 Zn2+ ions retained per dimer | Arabidopsis thaliana |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Arabidopsis thaliana | - |
AthTRZ1, wild-type and mutant variants studied | - |
Drosophila melanogaster | - |
- |
- |
Saccharomyces cerevisiae | P36159 | long tRNase Z enzyme | - |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
in vitro tRNA processing assays, bis(p-nitrophenyl)phosphate hydrolysis assay described, in contrast to the short tRNase Z enzyme AthTRZ1 of Arabidopsis thaliana, long tRNase Z enzymes do not have bis(p-nitrophenyl)phosphate hydrolysis activity | Saccharomyces cerevisiae |
additional information | - |
in vitro tRNA processing assays, bis(p-nitrophenyl)phosphate hydrolysis assay described, kinetic analysis shown, AthTRZ1 effective catalyst of bpNPP hydrolysis, AthTRZ1-R252G reveals ten times higher activity compared to the wild-type enzyme, in contrast to the short tRNase Z enzyme AthTRZ1, long tRNase Z enzymes do not have bis(p-nitrophenyl)phosphate hydrolysis activity | Arabidopsis thaliana |
additional information | - |
in vitro tRNA processing assays, bpNPP hydrolysis assay described, in contrast to the short tRNase Z enzyme AthTRZ1 of Arabidopsis thaliana, long tRNase Z enzymes do not have bis(p-nitrophenyl)phosphate hydrolysis activity | Drosophila melanogaster |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
bis(p-nitrophenyl)phosphate + H2O | differentiation of tRNase Z variants by 3'-processing activity and their ability to hydrolyze the phosphodiester bond in the chromogenic phosphodiester bis(p-nitrophenyl)phosphate (bpNPP), smallest known tRNase Z substrate, fourteen variants lost ability to hydrolyze bpNPP, seven variants reveal reduced activity | Arabidopsis thaliana | p-nitrophenol + p-nitrophenyl phosphate | - |
? | |
bis(p-nitrophenyl)phosphate + H2O | no hydrolysis of the the chromogenic phosphodiester bis(p-nitrophenyl)phosphate (bpNPP) observed | Drosophila melanogaster | p-nitrophenol + p-nitrophenyl phosphate | - |
? | |
bis(p-nitrophenyl)phosphate + H2O | no hydrolysis of the the chromogenic phosphodiester bis(p-nitrophenyl)phosphate (bpNPP) observed | Saccharomyces cerevisiae | p-nitrophenol + p-nitrophenyl phosphate | - |
? |
Subunits | Comment | Organism |
---|---|---|
dimer | - |
Arabidopsis thaliana |
Synonyms | Comment | Organism |
---|---|---|
AthTrz1 | - |
Arabidopsis thaliana |
tRNase Z | - |
Drosophila melanogaster |
tRNase Z | - |
Arabidopsis thaliana |
tRNase Z | - |
Saccharomyces cerevisiae |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | turnover rate of AthTRZ1 eight-fold reduced if compared to tRNAse Z of Escherichia coli | Arabidopsis thaliana |