Electrophoretic analysis of erythrocyte membrane proteins and glycoproteins from different species
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The erythrocyte membrane proteins and glycoproteins of man, rat, mouse, sheep and dog were analysed by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE), using a discontinuous buffer system. Considerable similarities between the species were observed in the pattern of protein bands seen when gels were stained with Coomassie Blue. Equivalents to human Bands 1, 2, 3, 4.2, 5 and 8 appeared to be present in the rat, mouse, sheep and dog, and Band 4.1 was identified as a closely spaced doublet in all species except the rat.
When RBC membranes were stained with periodic acid-Schiff (PAS) after SDS-PAGE analysis, glycoproteins equivalent to human glycophorins were identified in all the species studied. However, in contrast to the overall similarity of the protein patterns, the number, relative staining intensity and apparent molecular masses of the PAS-stained bands differed between species.
The silver stain was assessed in the detection of RBC membrane proteins in polyacrylamide gels, and found to be more sensitive than Coomassie Blue. The technique also stained many of the PAS-positive glycophorins as diffuse orange zones, which could be distinguished from the darker protein bands by their differential colouration.
In view of the interspecies variation in the glycophorins after SDS-PAGE, it is suggested that, unlike the membrane proteins, their functions do not require a conserved structure.
KeywordsErythrocyte Membrane proteins Membrane glycoproteins Comparative study Gel electrophoresis
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