Purification, renaturation, and reconstituted protein kinase activity of the Sendai virus large (L) protein: L protein phosphorylates the NP and P proteins in vitro.
Sodium dodecyl sulfate-solubilized Sendai virus large (L) protein was highly purified by a one-step procedure, using hydroxylapatite column chromatography. Monoclonal antibodies addressed to the carboxyl-terminal amino acid sequence of the L protein were used for monitoring L protein during purification. By removing sodium dodecyl sulfate from purified L protein, a protein kinase activity was successfully renatured. P and NP proteins served as its substrates. After immunoprecipitation with anti-L antibodies, the immunocomplex already showed protein kinase activity. In the presence of P protein, the NP protein was more highly phosphorylated. The results show that Sendai virus L protein possesses a protein kinase activity phosphorylating the other proteins of the viral nucleocapsid in vitro.
Einberger
H
H
Abteilung für Virusforschung, Max-Planck-Institut für Biochemie, Martinsried, Federal Republic of Germany.
Mertz
R
R
Hofschneider
P H
PH
Neubert
W J
WJ
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D015153:Blotting, Western; D002213:Capsid / Q000378:metabolism; D002642:Chick Embryo; D002846:Chromatography, Affinity; D012321:DNA-Directed RNA Polymerases; D004591:Electrophoresis, Polyacrylamide Gel; D007700:Kinetics; D008969:Molecular Sequence Data; D019590:Nucleocapsid Proteins; D009698:Nucleoproteins; D009842:Oligopeptides / Q000138:chemical synthesis; D010222:Parainfluenza Virus 1, Human / Q000201:enzymology*; D010750:Phosphoproteins / Q000378:metabolism*; D011489:Protein Denaturation; D011494:Protein Kinases / Q000302:isolation & purification / Q000378:metabolism*; D014758:Viral Core Proteins / Q000378:metabolism*; D014764:Viral Proteins / Q000302:isolation & purification / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D019590:Nucleocapsid Proteins; D009698:Nucleoproteins; D009842:Oligopeptides; C055859:P protein, Sendai virus; D010750:Phosphoproteins; D014758:Viral Core Proteins; D014764:Viral Proteins; D011494:Protein Kinases; C064365:L protein, Sendai virus; D012321:DNA-Directed RNA Polymerases