Lactoperoxidase-catalysed iodination of surface proteins on human melanoma cells

Abstract

The cell-surface proteins of 6 different melanoma cell cultures have been labelled with 125I using lactaperoxidase-catalysed iodination. Fractionation of the proteins was achieved using 5--22.5% polacrylamide-gradient gel electrophoresis in the presence of sodium dodecyl sulphate (SDS) and the proteins were detected by autoradiography. Up to 24 labelled proteins were detected in the individual cell cultures, but the proteins labelled differed considerably in the 6 cultures examined. A possible reason for this, involving variation in the glycosylation of cell-surface glycoproteins is discussed. Cells of the same melanoma line had similar cell-surface proteins at different passage levels, but changes in the labelled proteins occurred when the culture conditions were altered. The cell-surface proteins of high molecular weight were cleaved by trypsin, but most of the low mol.-wt. proteins were resistant to trypsin. The "large external transformation sensitive" (LETS) protein detected as a major protein on fibroblasts in culture was not a dominant protein on the melanoma cells. It was detected on only 4/6 cell cultures. Possible relationships of the cell-surface proteins described in this study to morphology, immunological properties and proteolytic activity of human melanoma cells are discussed.