IMMUNOELECTROPHORETIC PATTERN OF NATIVE MUCOSAL INTRACELLULAR GLYCOPROTEINS OF HOG HEALTHY AND DRUG-INTOXICATED STOMACHS AND OF HOG BODY-FLUIDS

Naturally occurring glycoproteins have been extracted from fundic and antral mucosal tissue of the hog stomach by means of nondegrading tech niques. Major and retarded glycoprotein fractions separated by gel fil tration were further dissociated from appreciable amounts of noncovale ntly bound proteins by CsCl density gradient centrifugation. Antisera to glycoprotein fractions of fundic and antral regions of the stomach were prepared in rabbits. The major fractions from both gastric region s have similar molecular mass (approximately 2 x 10(6)), sedimentation coefficient (approximately 31.5 s), and specific viscosity (approxima tely 1.6). Purified fractions from each region were further separated into two subfractions by affinity chromatography on wheat germ lectin. Glycoprotein subfractions from antrum and fundus differ appreciably i n their carbohydrate and amino acids content, share antigenic determin ants, but do not cross-react with anti-hog serum protein antisera. Fur ther diversity in native mucin glycoproteins was observed by the use o f one-(D) and two-dimensional (2D) immunoelectrophoresis; subfractions that cross-react with specific anti-hog gastric glycoproteins were fo und to contain three or more components. D-Immunoelectrophoretic analy ses demonstrated (1) in vivo degradation of glycoprotein components of the major fundic fraction isolated from mucosal tissue of alcohol/ace tyl salicylate-intoxicated hog stomachs and (2) in vitro catabolism of major fundic glycoproteins by corresponding mitochondrial lysosomal ( ML) acid hydrolases. Furthermore, 2D-immunoelectrophoretic analyses sh owed that (1) hog synovial fluid and plasma proteins have similar pros thetic moities as either reacted with anti-hog serum proteins antisera . Nonetheless, locations, shapes, and staining intensities of the immu noprecipitate lines differed, which is indicative of different structu res of the carbohydrate moities of components of synovial fluid and pl asma proteins, and (2) only a minor fraction of hog cerebrosplenal flu id cross-reacted with anti-hog serum protein antisera. This is contrar y to the generally accepted deduction based high-resolution 2D-electro phoresis, indicative of different compositional patterns of plasma and cerehrosplnal fluids.