title: Biochemical studies of the excitable membrane of Paramecium tetraurelia. V. Effects of proteases on the ciliary membrane.
abstract: The swimming behavior of Paramecium is regulated by an excitable membrane that covers the body and cilia of the protozoan. In order to obtain information on the topology and function of ciliary membrane proteins, Paramecia were treated with trypsin, chymotrypsin or pronase and the effects of these proteases were analyzed using electron microscopy, gel electrophoresis of ciliary fractions and behavioral tests. At the concentrations used, trypsin and chymotrypsin had little or no effect on the cells while pronase removed the cell surface coat, visible as fuzzy material covering the cell membrane. The same pronase treatment caused the specific removal of a high molecular weight protein (250 000), as judged by sodium dodecyl sulfate polyacrylamide gel electrophoresis. This protein, the 'immobilization antigen', constitutes the major protein of the ciliary membrane. Although the immobilization antigen was removed (or markedly decreased), no marked and reproducible difference was observed in the swimming behavior of the treated cells. We also determined the effects of proteases on isolated ciliary fractions to explore the sidedness of ciliary membrane proteins. A set of proteins relatively resistant to protease digestion was identified; they may be intrinsic membrane proteins.
mesh_terms: D002462:Cell Membrane / Q000648:ultrastructure; D002918:Chymotrypsin / Q000378:metabolism; D002923:Cilia / Q000378:metabolism* / Q000648:ultrastructure; D004591:Electrophoresis, Polyacrylamide Gel; D008565:Membrane Proteins / Q000378:metabolism*; D008854:Microscopy, Electron; D008970:Molecular Weight; D009068:Movement; D010247:Paramecium / Q000378:metabolism*; D010447:Peptide Hydrolases / Q000378:metabolism*; D011402:Pronase / Q000378:metabolism; D014357:Trypsin / Q000378:metabolism
chemical_list: D008565:Membrane Proteins; D010447:Peptide Hydrolases; D002918:Chymotrypsin; D014357:Trypsin; D011402:Pronase