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Literature summary extracted from

  • Karkashon, S.; Hopkinson, A.; Levinger, L.
    tRNase Z catalysis and conserved residues on the carboxy side of the His cluster (2007), Biochemistry, 46, 9380-9387.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.1.26.11 baculovirus expression system, pFastbac HTA, transformation into insect cells Drosophila melanogaster

Protein Variants

EC Number Protein Variants Comment Organism
3.1.26.11 A631T HEAT-domain-variant Drosophila melanogaster
3.1.26.11 C584A motif I-variant Drosophila melanogaster
3.1.26.11 D610A motif IV-variant Drosophila melanogaster
3.1.26.11 E630A HEAT-domain-variant Drosophila melanogaster
3.1.26.11 F669A motif V-variant Drosophila melanogaster
3.1.26.11 G609A motif IV-variant Drosophila melanogaster
3.1.26.11 H583A motif I-variant Drosophila melanogaster
3.1.26.11 H629A HEAT-domain-variant Drosophila melanogaster
3.1.26.11 H646A HST loop-variant Drosophila melanogaster
3.1.26.11 H668A motif V-variant Drosophila melanogaster
3.1.26.11 K644A HST loop-variant Drosophila melanogaster
3.1.26.11 L643A HST loop-variant Drosophila melanogaster
3.1.26.11 M612A motif IV-variant Drosophila melanogaster
3.1.26.11 additional information Ala-scanning mutagenesis through five conserved loops on the carboxy side of motif II (motifs III, IV, HEAT,HST, and motif V) performed, pre-tRNA processing kinetics of the expressed variants studied Drosophila melanogaster
3.1.26.11 N586A motif I-variant Drosophila melanogaster
3.1.26.11 P585A motif I-variant Drosophila melanogaster
3.1.26.11 P613A motif IV-variant Drosophila melanogaster
3.1.26.11 Q651A HST loop-variant Drosophila melanogaster
3.1.26.11 Q671A motif V-variant Drosophila melanogaster
3.1.26.11 R582A motif I-variant Drosophila melanogaster
3.1.26.11 R642A HST loop-variant Drosophila melanogaster
3.1.26.11 R672A motif V-variant Drosophila melanogaster
3.1.26.11 S587A motif I-variant Drosophila melanogaster
3.1.26.11 S608A motif IV-variant Drosophila melanogaster
3.1.26.11 S647A HST loop-variant Drosophila melanogaster
3.1.26.11 S650A HST loop-variant Drosophila melanogaster
3.1.26.11 S670A motif V-variant Drosophila melanogaster
3.1.26.11 T611A motif IV-variant Drosophila melanogaster
3.1.26.11 T632A HEAT-domain-variant Drosophila melanogaster
3.1.26.11 T645A HST loop-variant Drosophila melanogaster
3.1.26.11 T648A HST loop-variant Drosophila melanogaster
3.1.26.11 T667A motif V-variant Drosophila melanogaster
3.1.26.11 V581A motif I-variant Drosophila melanogaster
3.1.26.11 V649A HST loop-variant Drosophila melanogaster
3.1.26.11 Y607A motif IV-variant Drosophila melanogaster
3.1.26.11 Y673A motif V-variant Drosophila melanogaster

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.1.26.11 additional information
-
 additional information kinetics of tRNase Z processing in wild-type and variants shown, effects of substitutions in conserved regions on the C-side of motif II indicated, KM-values for tested variants similar to the mean wild-type KM value of 15 nM, residues involved with metal ion binding and catalysis not directly involved with substrate recognition and binding Drosophila melanogaster

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.1.26.11 Zn2+ binding of two zinc ions coordinated by four residues in the signature His domain, catalytically important conserved contact between Glu231 and His24 shown Drosophila melanogaster

Organism

EC Number Organism UniProt Comment Textmining
3.1.26.11 Drosophila melanogaster Q8MKW7
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.1.26.11 nickel chelate affinity purification Drosophila melanogaster

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.1.26.11 additional information
-
 reaction kinetics of tRNase Z variants with Ala substitutions through conserved loops on the carboxy side of motif II shown, substitution of motif IV aspartate reduces catalytic efficiency more than 10000fold, Histidines in motif III, V, and the HST loop also functionally important, replacement of Glu in the HEAT loop with Ala reduces efficiency by about 1000fold Drosophila melanogaster

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.1.26.11 pre-tRNAarg + H2O the new substrate pre-tRNAArg reveals kinetic parameters with wild type tRNase Z similar to those reported for pre-tRNAHis, enzyme concentration of 25 pM Drosophila melanogaster ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.1.26.11 tRNase Z
-
 Drosophila melanogaster