Induction of surface glycoprotein expression by cyclic AMP in Chinese hamster ovary cells.

Surface expression of a membrane glycoprotein of 135,000 molecular weight (P135) was inducible by adenosine 3',5' -cyclic monophosphate in Chinese hamster ovary cells, CHO-K1 clone. Cells were cultured in the presence or absence of cyclic AMP derivatives, chemicals influencing cytoplasmic cyclic AMP levels, or inhibitors of protein or RNA synthesis. Surface proteins wre radiolabeled by a lactoperoxidase-catalyzed iodination reaction and analyzed by two-dimensional polyacrylamide gel electrophoresis. Surface expression of P135 increased 3---5-fold inthe presence of N6,O2' -dibutyryl cyclic AMP or 8-parachlorophenylthio cyclic AMP. Induction was also observed after treatment with prostaglandins E1 and F2 alpha, but not with sodium butyrate. Phosphodiesterase inhibitor, Roche compound Ro20-1724, enhanced the effect of N6,O2' -dibutyryl cyclic AMP. Metabolic incorporation of [35S]methionine into P135 was increased by N6,O2' -dibutyryl cyclic AMP. The induction was sensitive to inhibitors of protein and RNA biosynthesis. These results are consistent with a proposal that cyclic AMP controls the synthesis of this protein. Metabolic incorporation of a radioactive precursor suggested that P135 was a glucos-amine-containing glycoprotein. P135 appeared to be strongly associated with cell membrane because it was resistant to extraction of plasma membrane by cole 0.1 N NaOH.