Cloned (Comment) | Organism |
---|---|
overexpression of native and selenomethionine-containing wild-type protein subunit Ph1877p, and mutant protein subunits Ph1788p in Escherichia coli | Pyrococcus horikoshii |
Crystallization (Comment) | Organism |
---|---|
10 mg/ml purified recombinant selenomethionine labeled protein subunit Ph1877p, vapour diffusion against 22.5% PEG 6000 and 0.1 M HEPES, pH 7.5, hanging drop method, crystals are suspended on a loop in a thin liquid film of stabilizing solution and frozen directly for X-ray diffraction structure determination and analysis at 1.8 A resolution | Pyrococcus horikoshii |
Protein Variants | Comment | Organism |
---|---|---|
D180A | site-directed mutagenesis, activity of the holoenzyme reconstituted with the recombinant mutant protein subunit Ph1877p is unaltered compared to holoenzyme reconstituted with recombinant wild-type Ph1877p | Pyrococcus horikoshii |
D98A | site-directed mutagenesis, activity of the holoenzyme reconstituted with the recombinant mutant protein subunit Ph1877p is unaltered compared to holoenzyme reconstituted with recombinant wild-type Ph1877p | Pyrococcus horikoshii |
H114A | site-directed mutagenesis, activity of the holoenzyme reconstituted with the recombinant mutant protein subunit Ph1877p is unaltered compared to holoenzyme reconstituted with recombinant wild-type Ph1877p | Pyrococcus horikoshii |
K123A | site-directed mutagenesis, reconstitution of the holoenzyme with the recombinant mutant protein subunit Ph1877p results in reduced activity compared to holoenzyme reconstituted with recombinant wild-type Ph1877p | Pyrococcus horikoshii |
K158A | site-directed mutagenesis, activity of the holoenzyme reconstituted with the recombinant mutant protein subunit Ph1877p is unaltered compared to holoenzyme reconstituted with recombinant wild-type Ph1877p | Pyrococcus horikoshii |
K196A | site-directed mutagenesis, reconstitution of the holoenzyme with the recombinant mutant protein subunit Ph1877p results in reduced activity compared to holoenzyme reconstituted with recombinant wild-type Ph1877p | Pyrococcus horikoshii |
K42A | site-directed mutagenesis, activity of the holoenzyme reconstituted with the recombinant mutant protein subunit Ph1877p is unaltered compared to holoenzyme reconstituted with recombinant wild-type Ph1877p | Pyrococcus horikoshii |
R107A | site-directed mutagenesis, reconstitution of the holoenzyme with the recombinant mutant protein subunit Ph1877p results in reduced activity compared to holoenzyme reconstituted with recombinant wild-type Ph1877p | Pyrococcus horikoshii |
R176A | site-directed mutagenesis, reconstitution of the holoenzyme with the recombinant mutant protein subunit Ph1877p results in 78% reduced activity compared to holoenzyme reconstituted with recombinant wild-type Ph1877p | Pyrococcus horikoshii |
R68A | site-directed mutagenesis, reconstitution of the holoenzyme with the recombinant mutant protein subunit Ph1877p results in slightly reduced activity compared to holoenzyme reconstituted with recombinant wild-type Ph1877p | Pyrococcus horikoshii |
R87A | site-directed mutagenesis, activity of the holoenzyme reconstituted with the recombinant mutant protein subunit Ph1877p is unaltered compared to holoenzyme reconstituted with recombinant wild-type Ph1877p | Pyrococcus horikoshii |
R90A | site-directed mutagenesis, reconstitution of the holoenzyme with the recombinant mutant protein subunit Ph1877p results in reduced activity compared to holoenzyme reconstituted with recombinant wild-type Ph1877p | Pyrococcus horikoshii |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
tRNA precursor + H2O | Pyrococcus horikoshii | tRNA processing | mature tRNA + 5'-oligonucleotide | - |
? | |
tRNA precursor + H2O | Pyrococcus horikoshii OT-3 | tRNA processing | mature tRNA + 5'-oligonucleotide | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pyrococcus horikoshii | O59543 | strain OT3 | - |
Pyrococcus horikoshii OT-3 | O59543 | strain OT3 | - |
Posttranslational Modification | Comment | Organism |
---|---|---|
ribonucleoprotein | - |
Pyrococcus horikoshii |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant protein subunits from Escherichia coli | Pyrococcus horikoshii |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
endonucleolytic cleavage of RNA, removing 5'-extranucleotides from tRNA precursor | an RNA-containing enzyme, essential for tRNA processing, generates 5'-termini or mature tRNA molecules, amino acids residues Arg90, Arg107, Lys123, Arg176, and Lys196 are involved in interaction with enzyme RNA or with the pre-tRNA | Pyrococcus horikoshii |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
pre-tRNATyr + H2O | substrate from Pyrococcus horikoshii OT3, cleavage of 5'-terminal oligonucleotide | Pyrococcus horikoshii | mature tRNATyr + 5'-oligoribonucleotide | generates 5'-phosphate,3'-hydroxyl-product | ? | |
pre-tRNATyr + H2O | substrate from Pyrococcus horikoshii OT3, cleavage of 5'-terminal oligonucleotide | Pyrococcus horikoshii OT-3 | mature tRNATyr + 5'-oligoribonucleotide | generates 5'-phosphate,3'-hydroxyl-product | ? | |
tRNA precursor + H2O | tRNA processing | Pyrococcus horikoshii | mature tRNA + 5'-oligonucleotide | - |
? | |
tRNA precursor + H2O | tRNA processing | Pyrococcus horikoshii OT-3 | mature tRNA + 5'-oligonucleotide | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | protein Ph1877p is one of the essential protein components of the ribozyme and forms a TIM barrel structure consisting of 10 alpha-helices and 7 beta-strands, the protein shows a cluster of positively charged amino acid residues on the molecule surface | Pyrococcus horikoshii |
Synonyms | Comment | Organism |
---|---|---|
RNase P | - |
Pyrococcus horikoshii |