Britain Synthesis and Turnover of Plasma - Membrane Proteins and Glycoproteins in a Neuroblastoma Cell Line By ROGER

A kinetic analysis of the appearance of "4C-labelled proteins in the surface membranes isolated from exponentially growing neuroblastoma cells (N2a) showed that the total membrane proteins reached a steady-state specific radioactivity in 18-20h. However, examination of individual protein bands resolved by sodium dodecyl sulphate-ureapolyacrylamide-gel electrophoresis illustrated that differences in the kinetics of specific surface-membrane proteins could be detected. Although most of the protein bands reached a steady-state specific radioactivity at a time similar to that for total membrane proteins, at least two bands (mol.wt. 180000 and 130000) attained the steady-state within 8-10h. It was shown by the use of dual-labelling techniques that these two protein bands turned over in the surface membranes of neuroblastoma N2a cells at least 180 and 150% faster than the total membrane protein. These two proteins were glycosylated and located on the outer surface of the cells, since they were labelled with radioactive carbohydrates and readily removed by treatment of the intact neuroblastoma cell with proteinases.