The expression of canine cardiac phospholamban in heterologous systems.
A synthetic phospholamban gene has been cloned and expressed in Escherichia coli, producing both native phospholamban and a fusion protein with 81 amino acids of the influenza virus NS1 protein. Both the native phospholamban and fusion proteins produced extensive cell lysis upon induction of gene expression, but only the native protein underwent spontaneous pentamer formation in E. coli. Translation in vitro of mRNA produced by transcription in vitro of phospholamban cDNA was used to demonstrate the spontaneous aggregation of phospholamban to form pentamers in this system also, both in the presence and absence of exogenous microsomes from canine pancreas or heart. Phospholamban produced by translation in vitro was apparently susceptible to proteolysis by enzymes present in the particulate fractions in these experiments.
Cook
E A
EA
Department of Cellular Pharmacology, Smith Kline and French Research Ltd., Welwyn, Herts., U.K.
Huggins
J P
JP
Sathe
G
G
England
P J
PJ
Piggott
J R
JR
D000251:Adenosine Triphosphatases / Q000235:genetics*; D000818:Animals; D001483:Base Sequence; D002135:Calcium-Binding Proteins / Q000096:biosynthesis / Q000235:genetics*; D003001:Cloning, Molecular / Q000379:methods; D004285:Dogs; D004790:Enzyme Induction; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D015870:Gene Expression; D005796:Genes; D005813:Genes, Synthetic; D008969:Molecular Sequence Data; D010957:Plasmids; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D015183:Restriction Mapping; D014158:Transcription, Genetic; D000099211:Phospholamban
D000251:Adenosine Triphosphatases; D002135:Calcium-Binding Proteins; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D000099211:Phospholamban