Comparative determination of the particle weight of glycoproteins by SDS-PAGE and analytical ultracentrifugation

摘要: The particle weight of 21 glycoproteins has been determined by SDS-PAGE and high-speed sedimentation equilibrium, focussing on the influence carbohydrate content, partial specific volume (\(\bar v_2\)), relative molar mass (M r) results both techniques. Results were compared with literature data 22 nonconjugated proteins yielding following results: (i) \(\bar v_2\)of native decreases linearly increasing content. Therefore, v_2 - values\)of not accessible to experimental determination may be calculated. (ii) M r glycoprotein subunits shows anomalies in electrophoretic behaviour. Small amounts impurities have no result, if one refers only major band(s). Systematic variation parameters T C, as well use Ferguson plots leads insignificant improvements. There is correlation between content A possible explanation for this behaviour found different conformations sugar moiety, interactions electrophoresis gel, extent SDS binding, (iii) Analytical ultracentrifugation yields accurate values glycoproteins, independent size molecule. application technique, however, requires exact knowledge v_2\)and absence impurities, (iv) comparison subunit masses obtained two methods clearly proves superiority analytical ultracentrifugation, when SDS-PAGE.