Calcium dependency of the binding and mitogenicity of phytohemagglutinin: Differentiation between calcium-dependent and independent binding events

The same amount of phytohemagglutinin binds to lymphocytes at 37 °C and at 0 °C. The binding is inhibited by calcium chelators at 37 °C, but not at 0 °C, during the very first minutes of contact between lectin and cells. N-Acetylgalactosamine in high concentrations inhibits binding at both temperatures. The binding at 0 °C is abortive in the sense that it does not result in subsequent DNA synthesis. These findings indicate that the binding of phytohemagglutin (PHA) to the glycoprotein receptor on the cell surface does not in itself activate the immediate transducer of the mitogenic signal. The binding has to be accompanied by some change which involves calcium and occurs very rapidly, but cannot take place at 0 °C, to give a proper mitogenic anchorage of PHA to the cells.