A secreted form of the human interleukin 2 receptor encoded by an "anchor minus" cDNA

J Immunol. 1986 Jun 1;136(11):4099-105.

Abstract

The DNA sequence encoding all of the putative intracytoplasmic domain and most of the trans-membrane domain of the human IL 2 receptor was deleted from a full length receptor cDNA. After expression in mouse L cells, the resultant "anchor minus" cDNA was found to direct the synthesis of a secreted rather than membrane-associated form of the IL 2 receptor. The secreted receptor protein (44,000 to 46,000 Mr) retained the capacity to bind both IL 2 and the monoclonal anti-Tac antibody, as evidenced by retention on IL 2 and anti-Tac affinity columns, inhibition of [3H]-anti-Tac binding to HUT 102B2 cells, and partial inhibition of IL 2-induced CTLL proliferation. Removal of these domains from the IL 2 receptor did not alter the posttranslational processing or rate of export of the truncated receptor protein. These data confirm the proposed membrane orientation of the IL 2 receptor (NH2 terminus out, COOH terminus in) and underscore the anchoring function of this carboxy terminal receptor segment. The availability of such anchor minus receptor cDNA constructs may facilitate purification of large quantities of receptor protein for further analysis of receptor structure, valency, and localization of the IL 2 binding site(s).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Carbohydrate Conformation
  • Chromatography, Affinity
  • DNA / metabolism*
  • Interleukin-2 / genetics
  • Interleukin-2 / metabolism*
  • L Cells / metabolism
  • Membrane Proteins / genetics
  • Membrane Proteins / isolation & purification
  • Membrane Proteins / metabolism
  • Mice
  • Neuraminidase
  • Precipitin Tests
  • Protein Processing, Post-Translational
  • Receptors, Immunologic / genetics
  • Receptors, Immunologic / isolation & purification
  • Receptors, Immunologic / metabolism*
  • Receptors, Interleukin-2

Substances

  • Interleukin-2
  • Membrane Proteins
  • Receptors, Immunologic
  • Receptors, Interleukin-2
  • DNA
  • Neuraminidase