Summary
The localization of insulin on the surface of fat cell ghosts, incubated at physiological concentrations of the hormone, was tested with ferritin labelled insulin antibodies. Insulin binding sites, as revealed by this approach, were found to be uniformly distributed around ghost membranes. About 20% of the number of vesicles or 15% of the membrane surface area of a ghost preparation was tagged with ferritin. No ferritin labelling was observed when insulin was omitted from the reaction mixture or when it was replaced by proinsulin, insulin A and B chain, glucagon, ACTH or oxytocin. The membranes were also devoid of ferritin when the antiinsulin conjugate was substituted by unspecificγ-globulin conjugate. The insulin binding sites demonstrated by the immunoferritin technique resembled the insulin receptor of fat cell membranes in so far as they showed trypsin sensitivity, heat lability and temperature dependency of insulin dissociation. From the distance between the ferritin particles and the membrane surface it appears that the insulin binding sites are located very close to or within the ghost membrane. The number of ferritin particles on individual membranes per unit area was higher than the number of insulin receptors calculated from the data of insulin binding studies on the basis of equal distribution of the receptor sites.
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Supported by a Fellowship from the Alexander von Humboldt-Stiftung, Bad Godesberg, Germany.
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Siess, E.A., Nestorescu, M.L. & Wieland, O.H. Localization of insulin on fat cell ghosts by ferritin labelled insulin antibodies. Diabetologia 10, 439–447 (1974). https://doi.org/10.1007/BF01221635
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DOI: https://doi.org/10.1007/BF01221635