Production of bioactive salmon calcitonin from the nonendocrine cell lines COS-7 and CHO.
To produce bioactive salmon calcitonin from the conventional nonendocrine cell lines, COS-7 and CHO, we devised a salmon calcitonin expression vector by combining the amino-terminus of human calcitonin precursor with a salmon calcitonin sequence, inserting the efficient furin-cleavable processing sequence Arg-X-Arg-X-Lys-Arg before salmon calcitonin, and deleting the carboxyl-terminal extension peptide. This chimeric calcitonin precursor terminates at glycine to easily receive an amidation reaction. COS-7 and CHO produced a high level of bioactive calcitonin by the resorption pit formation assay. Although amidating activity is highly expressed in CHO, but only a little in COS-7 cells, both cells produced a similar level of bioactive calcitonin. Thus, the engineered salmon calcitonin expression vector enables nonendocrine cells even with low amidation activity to produce bioactive calcitonin.
Takahashi
K I
KI
Department of Molecular Medicine, Gunma University, Maebashi, Japan.
Liu
Y C
YC
Hayashi
N
N
Goto
F
F
Kato
M
M
Kawashima
H
H
Takeuchi
T
T
D000818:Animals; D001681:Biological Assay; D016466:CHO Cells; D019556:COS Cells; D002116:Calcitonin / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D002522:Chlorocebus aethiops; D006224:Cricetinae; D017077:Culture Media, Conditioned / Q000494:pharmacology; D003804:Dentin / Q000187:drug effects; D005822:Genetic Vectors; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D012474:Salmon; D014162:Transfection
D017077:Culture Media, Conditioned; D011993:Recombinant Fusion Proteins; D002116:Calcitonin