A re-examination of the effects of chymotrypsin and trypsin on the erythrocyte membrane surface topology.
Silver/Coomassie blue staining of human erythrocyte membrane electrophoretograms permits simultaneous visualization and color differentiation of asialoproteins, sialoglycoproteins and lipids in the same gel. Using this technique evidence is provided that chymotrypsin cleaves glycophorin A as well as band 3. The chymotryptic fragmentation pattern of glycophorin A in situ intact cells was different from that generated by trypsin treatment. Chymotryptic cleavage of band 3 generated two Coomassie blue stained fragments at 62,000 and 38,000 Mr, whereas simultaneous cleavage of glycophorin A dimer and glycophorin A B heterodimer yielded yellow silver stained fragments at 68,000 and 47,000 Mr. Trypsin cleaved glycophorin A dimer (88,000 Mr) and monomer (38,000 Mr) to form membrane associated fragments of Mr = 40,000 and 18,000 respectively.
D001457:Anion Exchange Protein 1, Erythrocyte / Q000032:analysis; D002918:Chymotrypsin / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D004910:Erythrocyte Membrane / Q000378:metabolism*; D004912:Erythrocytes / Q000378:metabolism; D006021:Glycophorins / Q000032:analysis; D006801:Humans; D008970:Molecular Weight; D009439:Neuraminidase / Q000378:metabolism; D012795:Sialoglycoproteins / Q000032:analysis; D013499:Surface Properties; D014357:Trypsin / Q000378:metabolism*
D001457:Anion Exchange Protein 1, Erythrocyte; D006021:Glycophorins; D012795:Sialoglycoproteins; D009439:Neuraminidase; D002918:Chymotrypsin; D014357:Trypsin