title: Production of bioactive salmon calcitonin from the nonendocrine cell lines COS-7 and CHO.
abstract: 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.
authors.affiliation: Department of Molecular Medicine, Gunma University, Maebashi, Japan.
authors.forename: K I
authors.initials: KI
authors.lastname: Takahashi
authors.forename: Y C
authors.initials: YC
authors.lastname: Liu
authors.forename: N
authors.initials: N
authors.lastname: Hayashi
authors.forename: F
authors.initials: F
authors.lastname: Goto
authors.forename: M
authors.initials: M
authors.lastname: Kato
authors.forename: H
authors.initials: H
authors.lastname: Kawashima
authors.forename: T
authors.initials: T
authors.lastname: Takeuchi
mesh_terms: 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
chemical_list: D017077:Culture Media, Conditioned; D011993:Recombinant Fusion Proteins; D002116:Calcitonin