Biosynthesis and hormone-regulated expression of secretory glycoproteins in rat liver and hepatoma cells. Effect of glucocorticoids and inflammation

• Published in: The Journal of biological chemistry Vol. 256; no. 19; pp. 10145 - 10155
• Main Authors: Baumann, H, Held, W A
• Format: Journal Article
• Language: English
• Published: United States American Society for Biochemistry and Molecular Biology 10.10.1981
• Subjects: Animals; Cell Membrane - metabolism; Dexamethasone - pharmacology; Glycoproteins - biosynthesis; Glycoproteins - genetics; Inflammation - metabolism; Liver - drug effects; Liver - metabolism; Liver Neoplasms, Experimental - metabolism; Molecular Weight; Pancreas - metabolism; Protein Biosynthesis - drug effects; Rats; Rats, Inbred BUF; RNA, Messenger - genetics
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/S0021-9258(19)68755-7

Exposure of rat hepatoma tissue culture cells to dexamethasone results in appearance of a new glycoprotein, gp35-50 (Mr = 35,000 to 50,000) and increased synthesis of another glycoprotein, gp50 (Mr = 50,000). The glycoproteins synthesized in a fractionated cell-free translation system (containing dog pancreas microsomes) appear as a well separated series of spots on two-dimensional polyacrylamide gels which differ in size and charge. Glycoproteins synthesized by glucose-starved cells show similar size and charge heterogeneity. The size heterogeneity consists of a series of spots, each differing in molecular weights by about 3,000, which could be almost completely abolished by treatment with endo-beta-N-acetylglucosaminidase H. Our results indicate that unglycosylated gp50 (Mr = 42,000) typically acquires 3 N-glycan units, whereas gp35-50 (Mr = 22,000) possesses eight N-glycosylation sites. Analysis of the cell-free translation products directed by mRNA from hepatoma tissue culture cells grown in tissue culture, from hepatoma tissue, and from normal liver tissue indicated that administration of dexamethasone causes a pronounced increase in gp35-50 mRNA in all three tissues. A similar increase was observed in liver after inflammation which along with other biochemical properties suggests that gp35-50 may be alpha 1-acid protein. In contrast, mRNA coding for gp50 was not increased by dexamethasone in tumor tissue and no protein structurally related to gp50 was detected in the liver mRNA translation products. Thus, gp35-50 is expressed in normal liver, whereas gp50 is expressed in hepatoma cells and is differentially regulated by steroid hormones depending on whether the cells are grown in tissue culture or as a tumor in the rat.