STUDIES ON PROTEIN UPTAKE BY ISOLATED TUMOR CELLS

Abstract

Surface adsorption is studied in some detail because it is believed to be a major artifact in measurements of protein uptake by mammalian cells. Adsorption increases linearly with the I131-albumin concentration between 0.001 and 300 mg/ml. After short exposure to 300 mg/ml and two cell washings, the adsorption amounts to 38 mg albumin per gm cell proteins. Further washings remove 80 per cent of this value, leaving a small irreversibly bound residue. At equilibrium, adsorbed albumin can be labeled by a simple albumin exchange. This labeling reaches a steady state within seconds and stays at constant level over 30 minutes. Significant increases above this initial level are measured over periods of 2 hours. In our experimental conditions these increases can be considered due to albumin uptake. This uptake rises linearly with the albumin concentration between 0.5 and 50.0 mg/ml, and reaches 0.2 mg/gm cell protein or 4 x 105 molecules per cell. Compared to the incorporation of free amino acids in similar conditions, this value does not appear to contribute significantly to the N-metabolism of the tumor cells. Adsorption was generally greater than uptake. Both processes are linear functions of the same variable over the whole range of concentration tested. It is suggested that albumin is taken up by pinocytosis.