PubMed Health⌕ Search

Biomedical subjects

M Imada

Publications and source records attributed to M Imada.

66 records · Page 4Linked to original sources
PubMed1980

A two-dimensional polyacrylamide gel electrophoresis system for the analysis of mammalian cell surface proteins.

A detailed analysis of mammalian cell surface proteins is described by a new two-dimensional polyacrylamide gel electrophoresis technique. The first dimension gel contains 2% acrylamide, 0.1% sodium dodecyl sulfate, 0.3% Triton CF10 and 9 M urea. A combination of the detergents and urea permits the separation of poorly soluble, hydrophobic cell surface proteins. Under these conditions, the molecular size of proteins has a limited contribution to the fianl separation due to a low acrylamide concentration. Differences in charge properties, hydrophobicity, and glycosylation are the elements determining the resolution. In the second dimension, the proteins are separated primarily according to molecular weights, by a conventional polyacrylamide gel system in the presence of 0.1% sodium dodecyl sulfate. In this study, proteins of C6 rat glioma cell line are characterized. Cell surface proteins are specifically radio-labeled with 125I by a lactoperoxidase method, and compared with presumptive integral surface proteins which are resistant to extraction with 0.1 M NaOH. Also studied are total cellular proteins, fucose- and glucosamine-containing glycoproteins, and protein species with variable susceptibility to weak trypsin digestion. The electrophoresis system allows an unambiguous identification of each protein species.

Animals↗
PubMed1980

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.

4-(3-Butoxy-4-methoxybenzyl)-2-imidazolidinone↗