Characterization of insulin uptake into subcellular fractions of perfused rat liver using two different iodinated tracers

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Abstract
The binding and uptake of insulin in perfused rat liver has been investigated with specifically labelled 125I‐A14‐tyrosyl insulin as a tracer and compared with a commercially available iodo‐insulin preparation. The commercial preparation did not show saturation uptake kinetics and the clearence from the perfusate remained low and constant throughtout a wide concentration range. A14 labelled insulin showed saturation kinetics and high clearence at low carrier concentration, falling rapidly with increasing carrier concentration and reaching a steady state value of 1 ml/min. These results emphasize the importance of using specifically labelled insulin in physiological and biochemical studies of hepatic insulin metabolism. Perfusion with A14 tyrosine‐labelled insulin at 4°C showed apparent saturation with binding to the plasma membrane fraction. Perfusion at 37°C also showed apparent saturation with uptake predominantly to the ligandosome fraction. These results implicate the plasma membrane‐ligandosome pathway in the hepatic uptake of insulin at both physiological and pharmacological concentrations of the hormone.