MHC I Expression Regulates Co-clustering and Mobility of Interleukin-2 and -15 Receptors in T Cells

Gábor Mocsár, Julianna Volkó, Daniel Rönnlund, Jerker Widengren, Péter Nagy, János Szöllősi, Katalin Tóth, Carolyn K. Goldman, Sándor Damjanovich, Thomas A. Waldmann, Andrea Bodnár, György Vámosi

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Abstract

MHC glycoproteins form supramolecular clusters with interleukin-2 and -15 receptors in lipid rafts of T cells. The role of highly expressed MHC I in maintaining these clusters is unknown. We knocked down MHC I in FT7.10 human T cells, and studied protein clustering at two hierarchic levels: molecular aggregations and mobility by Förster resonance energy transfer and fluorescence correlation spectroscopy; and segregation into larger domains or superclusters by superresolution stimulated emission depletion microscopy. Fluorescence correlation spectroscopy-based molecular brightness analysis revealed that the studied molecules diffused as tight aggregates of several proteins of a kind. Knockdown reduced the number of MHC I containing molecular aggregates and their average MHC I content, and decreased the heteroassociation of MHC I with IL-2Rα/IL-15Rα. The mobility of not only MHC I but also that of IL-2Rα/IL-15Rα increased, corroborating the general size decrease of tight aggregates. A multifaceted analysis of stimulated emission depletion images revealed that the diameter of MHC I superclusters diminished from 400–600 to 200–300 nm, whereas those of IL-2Rα/IL-15Rα hardly changed. MHC I and IL-2Rα/IL-15Rα colocalized with GM1 ganglioside-rich lipid rafts, but MHC I clusters retracted to smaller subsets of GM1- and IL-2Rα/IL-15Rα-rich areas upon knockdown. Our results prove that changes in expression level may significantly alter the organization and mobility of interacting membrane proteins.

Original languageEnglish
Pages (from-to)100-112
Number of pages13
JournalBiophysical journal
Volume111
Issue number1
DOIs
Publication statusPublished - Jul 12 2016

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ASJC Scopus subject areas

  • Biophysics

Cite this

Mocsár, G., Volkó, J., Rönnlund, D., Widengren, J., Nagy, P., Szöllősi, J., Tóth, K., Goldman, C. K., Damjanovich, S., Waldmann, T. A., Bodnár, A., & Vámosi, G. (2016). MHC I Expression Regulates Co-clustering and Mobility of Interleukin-2 and -15 Receptors in T Cells. Biophysical journal, 111(1), 100-112. https://doi.org/10.1016/j.bpj.2016.05.044