Synthesis and release of glycosylated prolactin in transfected cells with human prolactin complementary deoxyribonucleic acid.
To analyze how the synthesis and release of glycosylated PRL (G-PRL) is regulated, we transfected human PRL complementary deoxyribonucleic acid (cDNA) into three different cell lines consisting of GH3 cells that originated in rat pituitary tissue, Chinese hamster ovary cells, and COS-1 cells generated from monkey renal tissue. 35S-labeled PRLs produced by the cells were immunoprecipitated with anti-human PRL antiserum, and the ratios of G-PRL to total PRL were compared. PRLs of 23 kDa and 25 kDa were detected in the cell lysate and medium. The 25-kDa PRL was confirmed to be a glycosylated form by endoglycosidase treatments. The ratios of G-PRL/total PRL were 0.17-0.33, which were similar in lysates and media and among different cell lines. Pulse-chase experiments revealed that the autonomaous secretion rates of G-PRL and non-glycosylated PRL were almost identical. These results indicate that synthesis and secretion kinetics of human PRL may not be affected by its glycosylation in the cells transfected with PRL cDNA.
Hirooka
T
T
Department of Obstetrics and Gynecology, Nagoya University School of Medicine, Japan.
Suganuma
N
N
Furuhashi
M
M
Kitagawa
T
T
Ando
H
H
Kikkawa
F
F
Tomoda
Y
Y
Seo
H
H
D000818:Animals; D016466:CHO Cells; D019556:COS Cells; D002460:Cell Line; D006224:Cricetinae; D018076:DNA, Complementary / Q000235:genetics*; D006026:Glycoside Hydrolases / Q000494:pharmacology; D006801:Humans; D007163:Immunosorbent Techniques; D007700:Kinetics; D008970:Molecular Weight; D010902:Pituitary Gland; D011388:Prolactin / Q000031:analogs & derivatives* / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D051381:Rats; D014162:Transfection
D018076:DNA, Complementary; C043928:prolactin, glycosylated; D011388:Prolactin; D006026:Glycoside Hydrolases