Establishment of the integrity of lysosomes in a glycoprotein-rich matrix. Distribution pattern of seven lysosomal enzymes in gastric mucosa.
Gel-forming mucosal glycoproteins strongly interfere with standard methods of cell fractionation. Thus, acid hydrolase-bound particles imbedded in the gel, sediment on centrifugation, in the nuclear fraction of homogenates of canine antral mucosa. These particles can be cleared by direct solubilization of the gel; however, the viscosity of the solution obtained prevents sedimentation of some of the latent hydrolases, even at very high speeds. The use of a new step-wise scheme of centrifugation and dilution successfully isolates lysosomal particles containing acid hydrolases from mucin-rich mucosa. All of the enzymes investigated, including acid phosphatase, cathepsin D, alpha- and beta-galactosidase, beta-B-acetylhexosaminidases, but with the exception of alpha-fucosidase, were found to be particle bound, exhibiting high degrees of latency. However, active mucosal particles are polydisperase in size and density, sedimenting under different centrifugal forces.
D000118:Acetylglucosaminidase / Q000032:analysis; D000135:Acid Phosphatase / Q000032:analysis; D000818:Animals; D002403:Cathepsins / Q000032:analysis; D002458:Cell Fractionation / Q000379:methods; D004285:Dogs; D005696:Galactosidases / Q000032:analysis; D005753:Gastric Mucosa / Q000201:enzymology*; D006596:Hexosaminidases / Q000032:analysis; D006867:Hydrolases / Q000032:analysis*; D008099:Liver / Q000201:enzymology; D008247:Lysosomes / Q000201:enzymology*; D008297:Male; D013385:Succinate Dehydrogenase / Q000032:analysis; D014461:Ultracentrifugation; D005644:alpha-L-Fucosidase / Q000032:analysis
D013385:Succinate Dehydrogenase; D006867:Hydrolases; D000135:Acid Phosphatase; D005696:Galactosidases; D006596:Hexosaminidases; D005644:alpha-L-Fucosidase; D000118:Acetylglucosaminidase; D002403:Cathepsins