PARTIAL-PURIFICATION AND CHARACTERIZATION OF RNASE-P FROM DICTYOSTELIUM-DISCOIDEUM

被引：0
作者：
STATHOPOULOS, C
KALPAXIS, DL
DRAINAS, D
机构：
来源：
EUROPEAN JOURNAL OF BIOCHEMISTRY | 1995年 / 228卷 / 03期
关键词：
DICTYOSTELIUM DISCOIDEUM; PURIFICATION; RIBONUCLEASE P; RIBOZYME;
D O I：
10.1111/j.1432-1033.1995.0976m.x
中图分类号：
Q5 [生物化学]; Q7 [分子生物学];
学科分类号：
071010 ; 081704 ;
摘要：
Ribonuclease P (RNase P) from Dictyostelium discoideum has been purified 470-fold. D. discoideum RNase P cleaves the precursor to Schizosaccharomyces pombe suppressor tRNA(Ser) at the same site as S. pombe RNase P, producing the mature 5' end of tRNA(Ser). pH and temperature optima for enzyme activity are 7.6 and 37 degrees C, respectively. The enzyme shows optimal activity in the presence of 5 mM MgCl2 and 10 mM NH4Cl or 5 mM KCl. The apparent K-m for the S. pombe tRNA precursor derived from the supSl tRNA(Ser) gene is 240 nM, and the apparent V-max is 3.6 pmol/min. Inhibition of D. discoideum RNase P by proteinase K and micrococcal nuclease strongly indicates that the activity requires both protein and RNA components. In cesium sulfate density gradients, the enzyme has a buoyant density of 1.23 g/ml, indicating a low RNA/protein ratio for the holoenzyme.
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页码：976 / 980
页数：5
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