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Molecular changes in cell surface membranes resulting from trypsinization of sarcoma 180 tumor cells
Authors:John W Huggins  Robert W Chestnut  Norman N Durham  Kermit L Carraway
Institution:1. Departments of Biochemistry, Oklahoma State University, Stillwater, Okla 74074 U.S.A.;2. Departments of Microbiology, Oklahoma State University, Stillwater, Okla 74074 U.S.A.
Abstract:Sarcoma-180 tumor cells in culture or grown as an ascites form in the CD-1 mouse have been subjected to mild trypsinization procedures in order to study morphological and molecular changes resulting from proteolysis. The cells attached to a substratum become rounded within 20 min and most undergo cell division, but they do not detach from the substratum. Removal of trypsin permits the cells to go back to their original spindle shape over an 8–20 h period.Surface membranes were isolated from trypsinized ascites and cultured cells and subjected to dodecyl sulfate-acrylamide gel electrophoresis. Both cell types showed the same two kinds of changes in electrophoretic patterns. First, there was a loss of glycoproteins from both cell types, even though they show different complements of cell surface glycoproteins. Second, there is a loss of high molecular weight polypeptides, which have previously been suggested to play a role in membrane stabilization and cell shape. These results further implicate these polypeptides in the control of cell morphology and offer circumstantial evidence for transmembrane interactions of surface glycoproteins with the high molecular weight polypeptides as a factor in controlling cell morphology.
Keywords:199  medium 199 minus calf serum  S-180  sarcoma 180 tumor grown in cell culture  SA-180  sarcoma 180 tumor grown in vivo as ascites form  PMSF  phenylmethanesulfonyl flouride  dimenthylsulfoxide  HEPES
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