Isolation of high-affinity murine interleukin 2 receptors as detergent-resistant membrane complexes
|Published in||European Journal of Immunology. 1990, vol. 20, no. 7, p. 1497-1503|
|Abstract||Murine T cells and T cell lines bearing high- and low-affinity receptors for interleukin (IL) 2 were chemically cross-linked to radiolabeled IL 2 and subjected to differential detergent extractions to evaluate the extent of IL 2 receptor association with the nonionic detergent-resistant framework of the plasma membrane. Low-affinity receptors were readily solubilized by nonionic detergent extraction of whole cross-linked cells, while solubilization of high-affinity receptors required a stronger ionic detergent suggesting their association with a membrane structure that is resistant to nonionic detergents. To achieve physical separation of low- and high-affinity receptors, cells cross-linked to 125I-labeled IL 2 were centrifuged through a sucrose barrier containing Triton X-100. Alternatively, Triton X-114 extracts of plasma membrane fractions were partitioned into aqueous and detergent phases. By either approach, high-affinity receptors differed from low-affinity ones by their increased density and consisted of detergent-resistant complexes containing p55-p75 heterodimers. The low-affinity receptors, on the contrary, were of low density and consisted exclusively of detergent-soluble p55 subunits. High density and resistance to nonionic detergent extraction of high-affinity IL 2 receptors suggest their integration into lateral microdomains of the detergent-resistant framework of the plasma membrane.|
|Keywords||Animals — Cell Fractionation — Cell Membrane/metabolism — Centrifugation, Density Gradient — Cross-Linking Reagents — Detergents — Humans — Interleukin-2/metabolism — Molecular Weight — Octoxynol — Polyethylene Glycols — Receptors, Interleukin-2/ isolation & purification/metabolism — Sodium Dodecyl Sulfate — Solubility — Structure-Activity Relationship|
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|HOESSLI, Daniel, POINCELET, Monique, RUNGGER-BRANDLE, Elisabeth. Isolation of high-affinity murine interleukin 2 receptors as detergent-resistant membrane complexes. In: European Journal of Immunology, 1990, vol. 20, n° 7, p. 1497-1503. doi: 10.1002/eji.1830200714 https://archive-ouverte.unige.ch/unige:10910|