Crosslinking erythrocyte membranes

Reported applications of SADP include the crosslinking of con A to receptors oVn human erythrocyte membranes (Vanin and Ji, 1981), site-specific labeling of lectins using modified... 

Romer-lAithi, C.R., Hajdu, J., and Brodbeck, U. Molecular forms of purified human erythrocyte membrane acetylcholinesterase investigated by crosslinking with dlimldates. Hoppe-Seyler s Z. Physiol. Chem. 360 929-934, 1979. 

Fig. 5.6. Two-dimensional gel electrophoresis of reversibly crosslinked human erythrocyte membranes. Ghosts were crosslinked with methyl 3-(4-azidophenyldithio)propionimidate and electrophoresed on a 0.5% agarose-1.75% polyacrylamide tube gel. The gel was treated with P-mercaptoethanol (10 mM), and electrophoresed in the second dimension on a 4 to 12 % gradient polyacrylamide slab gel. Some of the crosslinked components (mostly oligomers of spectrin) were incompletely cleaved. (Ji et al., 1980).

While the two-dimensional electrophoretic analysis described above is suitable for biological complexes that contain only a few major components (such as the human erythrocyte membrane), it is not feasible to extend its use to more complex systems such as the plasma membranes of many other cell types. Nevertheless, several ways to improve the method may prove useful. Immunoprecipitation of all the crosslinking products... 

Stroma, erythrocyte membrane residues from haemolysed red cells, have been intra-crosslinked with glutaraldehyde and used for the affinity chromatographic purification of two nonspecific and noninhibitable lectins fi-om Phaseolus vulgaris and P. coccineus. Formaldehyde-fixed human erythrocytes have provided a specific adsorbent for the purification of the lectin from Robinia pseudocacia. ... 

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