Complete purification of the pseudorabies virus protein kinase.
The recently described pseudorabies virus protein kinase has been purified from infected hamster fibroblasts by a combination of anion-exchange, hydrophobic-interaction and affinity chromatography. The purification resulted in enzyme with a specific activity in excess of 1,000 nmol phosphate mg-1 min-1 in relatively high yield. Gel electrophoresis of the purified enzyme under denaturing conditions revealed a single stained band at a position of migration corresponding to a Mr 38,000. Incubation of the purified enzyme with [gamma-32P]ATP in the absence of added substrate resulted in incorporation of 32P into this protein band, consistent with the 38-kDa protein being a protein kinase with a capacity for autophosphorylation. The phosphorylated form of the protein has an isoelectric point of approximately 4.9. Gel permeation chromatography of the purified enzyme indicated a native Mr 70,000, suggesting that the protein kinase has a homodimeric structure.
Purves
F C
FC
Department of Biochemistry, University of Glasgow, Scotland.
Katan
M
M
Leader
D P
DP
D000818:Animals; D002460:Cell Line; D002846:Chromatography, Affinity; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D011558:Herpesvirus 1, Suid / Q000201:enzymology*; D046911:Macromolecular Substances; D008970:Molecular Weight; D011494:Protein Kinases / Q000302:isolation & purification* / Q000378:metabolism
D046911:Macromolecular Substances; D011494:Protein Kinases