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Isolation of high-affinity murine interleukin 2 receptors as detergent-resistant membrane complexes

Poincelet, Monique
Rungger-Brandle, Elisabeth
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 AnimalsCell FractionationCell Membrane/metabolismCentrifugation, Density GradientCross-Linking ReagentsDetergentsHumansInterleukin-2/metabolismMolecular WeightOctoxynolPolyethylene GlycolsReceptors, Interleukin-2/ isolation & purification/metabolismSodium Dodecyl SulfateSolubilityStructure-Activity Relationship
PMID: 2387314
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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

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Deposited on : 2010-08-26

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