Cloned (Comment) | Organism |
---|---|
baculovirus expression system, pFastbac HTA, transformation into insect cells | Drosophila melanogaster |
Protein Variants | Comment | Organism |
---|---|---|
A631T | HEAT-domain-variant | Drosophila melanogaster |
C584A | motif I-variant | Drosophila melanogaster |
D610A | motif IV-variant | Drosophila melanogaster |
E630A | HEAT-domain-variant | Drosophila melanogaster |
F669A | motif V-variant | Drosophila melanogaster |
G609A | motif IV-variant | Drosophila melanogaster |
H583A | motif I-variant | Drosophila melanogaster |
H629A | HEAT-domain-variant | Drosophila melanogaster |
H646A | HST loop-variant | Drosophila melanogaster |
H668A | motif V-variant | Drosophila melanogaster |
K644A | HST loop-variant | Drosophila melanogaster |
L643A | HST loop-variant | Drosophila melanogaster |
M612A | motif IV-variant | Drosophila melanogaster |
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 |
N586A | motif I-variant | Drosophila melanogaster |
P585A | motif I-variant | Drosophila melanogaster |
P613A | motif IV-variant | Drosophila melanogaster |
Q651A | HST loop-variant | Drosophila melanogaster |
Q671A | motif V-variant | Drosophila melanogaster |
R582A | motif I-variant | Drosophila melanogaster |
R642A | HST loop-variant | Drosophila melanogaster |
R672A | motif V-variant | Drosophila melanogaster |
S587A | motif I-variant | Drosophila melanogaster |
S608A | motif IV-variant | Drosophila melanogaster |
S647A | HST loop-variant | Drosophila melanogaster |
S650A | HST loop-variant | Drosophila melanogaster |
S670A | motif V-variant | Drosophila melanogaster |
T611A | motif IV-variant | Drosophila melanogaster |
T632A | HEAT-domain-variant | Drosophila melanogaster |
T645A | HST loop-variant | Drosophila melanogaster |
T648A | HST loop-variant | Drosophila melanogaster |
T667A | motif V-variant | Drosophila melanogaster |
V581A | motif I-variant | Drosophila melanogaster |
V649A | HST loop-variant | Drosophila melanogaster |
Y607A | motif IV-variant | Drosophila melanogaster |
Y673A | motif V-variant | Drosophila melanogaster |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
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 | Comment | Organism | Structure |
---|---|---|---|
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 | UniProt | Comment | Textmining |
---|---|---|---|
Drosophila melanogaster | Q8MKW7 | - |
- |
Purification (Comment) | Organism |
---|---|
nickel chelate affinity purification | Drosophila melanogaster |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
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 | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
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 | Comment | Organism |
---|---|---|
tRNase Z | - |
Drosophila melanogaster |