Isolation and analysis of an Abelson murine leukemia virus-encoded tyrosine-specific kinase produced in Escherichia coli.
A segment of the coding sequence of the Abelson murine leukemia virus transforming gene (v-abl) has been inserted into a plasmid vector that allows its efficient and regulated expression in Escherichia coli. The product of the v-abl-derived coding sequence, designated p60v-abl, accumulated to a level of approximately 10% of total E. coli protein. A procedure is described for the isolation of p60v-abl from E. coli that yields about 50 micrograms of p60v-abl/g wet weight of E. coli. p60v-abl was capable of autophosphorylation and phosphorylating certain E. coli proteins specifically at tyrosine residues. The E. coli-expressed p60v-abl specifically phosphorylated tyrosine residues on casein and angiotensin II. The Km and Vmax values for ATP, casein, and angiotensin II in the p60v-abl kinase reaction have been determined and compared to values reported for other tyrosine-specific kinases. The expression system and isolation procedure described here permit the preparation of functional p60v-abl in quantities sufficient for detailed physical and biochemical characterization and examination of its biological action(s).
Ferguson
B
B
Pritchard
M L
ML
Feild
J
J
Rieman
D
D
Greig
R G
RG
Poste
G
G
Rosenberg
M
M
D000011:Abelson murine leukemia virus / Q000201:enzymology / Q000235:genetics*; D000804:Angiotensin II / Q000378:metabolism; D002364:Caseins / Q000378:metabolism; D003001:Cloning, Molecular; D004262:DNA Restriction Enzymes / Q000378:metabolism; D004279:DNA, Viral / Q000032:analysis; D004926:Escherichia coli / Q000378:metabolism*; D005786:Gene Expression Regulation; D009052:Leukemia Virus, Murine / Q000235:genetics*; D010957:Plasmids; D011494:Protein Kinases / Q000235:genetics*; D011505:Protein-Tyrosine Kinases; D014162:Transfection
D002364:Caseins; D004279:DNA, Viral; D000804:Angiotensin II; D011494:Protein Kinases; D011505:Protein-Tyrosine Kinases; D004262:DNA Restriction Enzymes