Expression and characterization of the tau subunit of phosphorylase kinase.
A cDNA encoding the entire tau subunit of rabbit skeletal muscle phosphorylase kinase was reconstructed and inserted into a plasmid containing the Escherichia coli ptac promoter and a constructed plasmid containing the ptac promoter and bacterial chloramphenicol acetyl transferase (CAT) gene, respectively. A significant phosphorylase kinase activity was found, in the first case. In the second case, a fused protein containing 73 amino acids from the CAT protein was obtained. After renaturation, the CAT-tau subunit protein shows enzymatic activity similar to the HPLC-purified and renatured tau subunit.
Chen
L R
LR
Dept. of Biochem. and Biophys., Iowa State Univ., Ames 50011.
Yuan
C J
CJ
Somasekhar
G
G
Wejksnora
P
P
Peterson
J E
JE
Myers
A M
AM
Graves
L
L
Cohen
P T
PT
da Cruz e Silva
E F
EF
Graves
D J
DJ
D000818:Animals; D015153:Blotting, Western; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D007700:Kinetics; D046911:Macromolecular Substances; D010764:Phosphorylase Kinase / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013329:Structure-Activity Relationship
D046911:Macromolecular Substances; D011993:Recombinant Fusion Proteins; D004247:DNA; D010764:Phosphorylase Kinase