Species variability in the modification of erythrocyte surface proteins by enzymatic probes.
Bovine and equine erythrocytes have been studied by three different surface modification techniques to investigate the accessibility of the surface components to the external medium. Lactoperoxidase labeling of equine erythrocytes results in a significant labeling of only one membrane component, a 100 000-mol.wt polypeptide corresponding to the membrane-spanning Component III of human erythrocytes. The major sialoglycoprotein of the equine erythrocyte is not labeled. This is in contradistinction to the situation for human and bovine cells, where both components are labeled. The equine membrane sialoglycoprotein is also not markedly affected by pronase, chymotrypsin or trypsin treatment of whole cells under the treatment conditions used, although it can be cleaved by pronase in isolated membranes. Experiments with the isolated glycoprotein show that its cleavage by trypsin is quite selective, whereas cleavage by pronase and chymotrypsin is much more extensive. Labelling of bovine red cells by galactose oxidase treatment followed by reduction with 3H-labeled borohydride yields radioactivity in only one major peak, that corresponding increase in labeling. Equine erythrocytes don not show significant labeling by this technique unless a neuraminidase pretreatment has been performed. Then only the major glycoprotein is labeled. Thus the equine glycoprotein is apparently inaccessible to the cell surface by standard surface modification methods, although it is clearly a surface component. These experiments point out some of the limitations of surface labeling and proteolysis methods in probing the accessibility of membrane components. The results suggest that apparent inaccessibility of the equine glycoprotein is due partially to its structure and partially to its localization in the membrane.
D000429:Alcohol Oxidoreductases; D000818:Animals; D002417:Cattle; D002462:Cell Membrane / Q000201:enzymology*; D004912:Erythrocytes / Q000201:enzymology*; D005690:Galactose; D006023:Glycoproteins / Q000097:blood*; D006736:Horses; D007785:Lactose; D009439:Neuraminidase / Q000378:metabolism; D010544:Peroxidases; D011402:Pronase / Q000378:metabolism; D013045:Species Specificity; D014357:Trypsin / Q000378:metabolism
D006023:Glycoproteins; D000429:Alcohol Oxidoreductases; D010544:Peroxidases; D009439:Neuraminidase; D014357:Trypsin; D011402:Pronase; D007785:Lactose; D005690:Galactose