Limited Proteolysis in the Study of Membrane Proteins

摘要: A substantial proportion of glycoproteins mammalian cells is found associated with the surface membrane. Several modes association are known. For example, transmembrane or integral proteins linked membrane by specific stretches protein consisting predominantly hydrophobic amino acids that function as anchors and span at least once. Some other covalently attached to a phospholipid in through an oligosaccharide (denoted glycophosphatidyl inositol-(GPI)-anchored proteins). Association cell even types these associations could be determined treatment intact vesicular membranes limited dilutions proteolytic enzymes, such trypsin. expression recombinant pro-lactase-phlorizin hydrolase (pro-LPH) transfected COS-1 (Nairn et al., 1991) hemagglutinin influenza virus (HA) infected CV-1 (Doyle 1985; Naim Roth, 1993) has been assessed susceptibilty The appearance specifically cleaved molecular species pro-LPH HA indicative for presence molecules surface.