标题:Biochemical Characterization of 60S Acidic Ribosomal P Proteins Associated with CK-II from Bamboo Shoots and Potent Inhibitors of Their Phosphorylation in Vitro
摘要:Three effective phosphate acceptors (35, 15 and 13 kDa polypeptides) for casein kinase II (CK-II) in the Superdex CK-II fraction prepared from a 0.5 M NaCl extract of bamboo shoots were selectively purified by glycyrrhizin (GL)-affinity column chromatography (HPLC). These three proteins (p35, p15 and p13) were identified as 60S acidic ribosomal P proteins by determination of their partial N-terminal sequences CK-II was associated with p35 since the GL-affinity fraction was coprecipitated with an anti-serum against Drosophila CK-IIβ. Moreover, a derivative (oGA) of glycyrrhetinic acid (GA) and several polyphenol-containing anti-oxidative compounds [quercetin, epigallocatechin gallate (FGCG) and two isoflavones, i.e., 3', 4', 7-trihydroxyisoflavone (3', 4', 7-THI) and 8-chloro-3', 4', 5, 7-tetrahydroxyisoflavone (8C-3', 4', 5, 7-THI)] inhibited the CK-II-mediated phosphorylation of 60S acidic ribosomal P proteins in vitro. Quercetin was found to be the most effective compounds on CK-II activity since its ID50 was approx. 50 nM. These results suggest that (i) GL-affinity column chromatography is useful for the selective purification of 60S acidic ribosomal P proteins as a heterocomplex associated with CK-II from various cell sources; (ii) natural anti-oxidative compounds with polyphenols, but not GL and GA, act as potent CK-II suppressors; and (iii) CK-II mediates the regulation of the physiological functions of 60S acidic ribosomal P proteins in growing plant cells.
关键词:60S acidic ribosomal P protein;casein kinase II;phosphorylation;CK-II inhibitor;glycyrrhizin;anti-oxidative compound