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pubmed-article:8086496pubmed:abstractTextA double-stranded DNA-dependent protein serine/threonine kinase (DNA-PK) was purified from a nuclear extract of Raji Burkitt's lymphoma cells by a three-step column-chromatographic procedure. The main silver-stained band visualized after SDS/PAGE corresponded to an autophosphorylated polypeptide of about 350-kDa that represents the catalytic component. The existence of Ku DNA-binding protein as a regulatory component in the purified enzyme was revealed by Western blot/enzyme immunoassay and direct inhibition test with anti-Ku sera from the autoimmune patients. The DNA-PK catalyzed phosphorylation of synthetic peptides corresponding to Myc and RB proteins in a DNA-dependent manner, indicating that DNA-PK may recognize a second core-sequence motif Pro-Ser/Thr- in addition to the putative consensus sequences of -Ser/Thr-Gln. The level of enzyme activity was significantly higher in DMSO-induced G0/G1-arrested Raji cells as well as in the cells after release from DMSO than in the log-phase cells.lld:pubmed
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pubmed-article:8086496pubmed:dateRevised2009-11-19lld:pubmed
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pubmed-article:8086496pubmed:articleTitleMolecular properties, substrate specificity and regulation of DNA-dependent protein kinase from Raji Burkitt's lymphoma cells.lld:pubmed
pubmed-article:8086496pubmed:affiliationDepartment of Pathological Biochemistry, Tokyo Medical and Dental University, Japan.lld:pubmed
pubmed-article:8086496pubmed:publicationTypeJournal Articlelld:pubmed
pubmed-article:8086496pubmed:publicationTypeResearch Support, Non-U.S. Gov'tlld:pubmed
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