[Couple production of human calcitonin and rat peptidylglycine alpha-amidation monooxygenase in insect cells].
Human calcitonin (hCT) is a 32 amino acid peptide hormone that requires C-terminal amidation for full biological activity. Calcitonin has important physiological function in vivo. We describe the couple expression of a synthesized modified human calcitonin(hmCT) gene fused with glutathione-S-transferase and rat peptidylglycine alpha-amidation monooxygenase (PAM) in insect cells infected by recombinant baculovirus GSTCT/PAM. Using Western blotting against hmCT or rat PAM, the GSThmCT fusion protein had been identified as well as the PAM. Following affinity chromatography with glutathione agarose column, the GSThmCT fusion protein produced by insect cells was purified. The purified fusion protein was also interacted with antibody against hmCT. The couple expression of a modification enzyme and its substrate in eucaryotic expression system may be used for producing other biological activity peptides.
Yang
Guan-Zhen
GZ
Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.
Chen
Zhen-Zhen
ZZ
Cui
Da-Fu
DF
Li
Bo-Liang
BL
Wu
Xiang-Fu
XF
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D002116:Calcitonin / Q000096:biosynthesis / Q000235:genetics*; D002478:Cells, Cultured; D015870:Gene Expression; D005982:Glutathione Transferase / Q000235:genetics*; D006801:Humans; D007313:Insecta / Q000166:cytology; D006899:Mixed Function Oxygenases / Q000096:biosynthesis / Q000235:genetics*; D009097:Multienzyme Complexes / Q000096:biosynthesis / Q000235:genetics*; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000235:genetics
D009097:Multienzyme Complexes; D011993:Recombinant Fusion Proteins; D002116:Calcitonin; D006899:Mixed Function Oxygenases; C035189:peptidylglycine monooxygenase; D005982:Glutathione Transferase