Expression of rat hepatic glucokinase in Escherichia coli.
Rat liver glucokinase was expressed in Escherichia coli by using an expression system based on bacteriophage T7 RNA polymerase. The expressed protein starts with the predicted initiator methionine residue and ends at the appropriate carboxyl terminal residue. It was partially purified by ammonium sulfate precipitation and gel filtration and had kinetic and physical properties similar to the purified rat liver enzyme. The efficient expression of this low abundance hepatic protein in bacteria provides a system for in vitro analysis of mutations of the enzyme.
Chien
C T
CT
Department of Physiology and Biophysics, State University of New York, Stony Brook 11794.
Tauler
A
A
Lange
A J
AJ
Chan
K
K
Printz
R L
RL
el-Maghrabi
M R
MR
Granner
D K
DK
Pilkis
S J
SJ
D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods*; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D015870:Gene Expression; D005796:Genes; D005822:Genetic Vectors; D005941:Glucokinase / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D008969:Molecular Sequence Data; D010957:Plasmids; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000378:metabolism; D015183:Restriction Mapping
D011994:Recombinant Proteins; D005941:Glucokinase