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The influence of non-specific binding on scatchard analysis used in insulin/receptor binding studies with erythrocytes
Authors:A Dwenger  P Tost and I Trautschold
Institution:(1) Abteilung für Klinische Biochemie des Zentrums Biochemie, Medizinische Hochschule Hannover, Karl-Wiechert-Allee 9, D-3000 Hannover 61, Germany
Abstract:Conclusions By equilibrium binding studies it could be demonstrated that erythrocytes have been saturated with specifically bound insulin in the order of 0.10–0.20 nmol/l with 4×1012 erythrocytes/l. From this an insulin receptor number of 15–30 receptors/normal erythrocyte could be calculated.By the current results we got evidence that in the insulin/erythrocyte receptor system the terminal part of the Scatchard plot was artefactual and might not be used for the calculation procedures because it is strongly influenced by non-specific effects. Because of the fluid change from the specific to the non-specific character of binding we propose to use total ligand concentrations only up to 5 nmol/l with 4×1012 erythrocytes/l for the investigation of specific binding on erythrocyte insulin receptors.By focussing on the initial part of the Scatchard plot curvilinearity disappeared and the character of this hormone/receptor system was reduced to one binding site with one characteristic affinity constant and a receptor concentration of one characteristic order.
Der Einfluß der unspezifischen Bindung bei Anwendung des Scatchard Plots zur Untersuchung der Insulin/Receptor-Wechselwirkungen an Erythrocyten
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