Characterization of the α subunit of the IFN-α receptor: Evidence of N- and O-linked glycosylation and association with other surface proteins

Leonidas C. Platanias, Lawrence M. Pfeffer, Ricardo Cruciani, Oscar R. Colamonici

Research output: Contribution to journalArticle

26 Scopus citations

Abstract

We studied the association of the α subunit of the (IFN-α-receptor) to other receptor components in the human H-929 and U-266 myeloma cell lines. Immunoprecipitation performed with the IFNaR3 mAb showed that two proteins with molecular masses of 205 and 145 kDa are co-precipitated with the α subunit. These complexes may not bind IFN-α as shown by studies using the heterobifunctional cross-linking reagent Denny-Jaffe and by partial cleavage of the homobifunctional cross-linker dithio succinimidyl propionate. These studies also provided evidence that at least two subunits with molecular mass of 130 kDa (α subunit) and 110 kDa (including 20 kDa corresponding to IFN- α) contribute to the formation of the IFN-α-receptor complex. To further characterize the α subunit of the IFN-α-receptor, immunoprecipitates using the mAb IFNaR3 were sequentially treated with N-glycanase, neuraminidase and O-glycanase. These studies showed that the α subunit is heavily glycosylated and has a protein precursor with a molecular mass of 68 kDa. Binding studies provided evidence for high and low affinity binding sites for IFN-α2. Affinity cross-linking experiments under low and high affinity conditions suggest that the high affinity binding site of the IFN-α-receptor is formed by a complex containing the α subunit, whereas the 110-kDa subunit may bind IFN-α2 under low affinity conditions.

Original languageEnglish (US)
Pages (from-to)3382-3388
Number of pages7
JournalJournal of Immunology
Volume150
Issue number8 I
StatePublished - Jan 1 1993

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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