Expression of human pituitary adenylate cyclase activating polypeptide (PACAP) cDNA in CHO cells and characterization of the products.
cDNA encoding human PACAP precursor was expressed in non-neuroendocrine Chinese hamster ovary cells, CHO-K1, The cells were transfected with expression vector (pTS705) containing the human PACAP cDNA by electroporation. A cell line which produced more than 80 ng/ml of immunoreactive PACAP (ir-PACAP) into the conditioned medium was established. RP-HPLC analysis of culture medium of this established cell line exhibited the presence of two types of PACAP, i.e. PACAP38 and PACAP27. At the same time, it was also revealed that immunoreactive PACAP-related peptide (ir-PRP) was secreted into the cultured medium. The ir-PACAPs were confirmed to ahve biological activities such as induction of cAMP and neurite outgrowth in rat pheochromocytoma PC12h cells.
Okazaki
K
K
Tsukuba Research Laboratories, Takeda Chemical Industries Ltd., Ibaraki, Japan.
Kimura
C
C
Kosaka
T
T
Watanabe
T
T
Ohkubo
S
S
Ogi
K
K
Kitada
C
C
Onda
H
H
Fujino
M
M
D000262:Adenylyl Cyclases / Q000378:metabolism*; D000818:Animals; D016466:CHO Cells / Q000201:enzymology*; D002461:Cell Line, Transformed; D002851:Chromatography, High Pressure Liquid; D006224:Cricetinae; D004247:DNA / Q000737:chemistry*; D005822:Genetic Vectors; D006801:Humans; D008970:Molecular Weight; D016501:Neurites / Q000737:chemistry; D009479:Neuropeptides / Q000235:genetics*; D051219:Pituitary Adenylate Cyclase-Activating Polypeptide; D010903:Pituitary Gland, Anterior / Q000201:enzymology*; D011498:Protein Precursors / Q000235:genetics / Q000276:immunology; D012333:RNA, Messenger / Q000737:chemistry; D051381:Rats
C495401:ADCYAP1 protein, human; C495403:Adcyap1 protein, rat; D009479:Neuropeptides; D051219:Pituitary Adenylate Cyclase-Activating Polypeptide; D011498:Protein Precursors; D012333:RNA, Messenger; D004247:DNA; D000262:Adenylyl Cyclases