CD20 homo-oligomers physically associate with the B cell antigen receptor. Dissociation upon receptor engagement and recruitment of phosphoproteins and calmodulin-binding proteins

J Biol Chem. 2008 Jul 4;283(27):18545-52. doi: 10.1074/jbc.M800784200. Epub 2008 May 12.

Abstract

B cell antigen receptor (BCR) signaling initiates sustained cellular calcium influx necessary for the development, differentiation, and activation of B lymphocytes. CD20 is a B cell-restricted tetraspanning protein organized in the plasma membrane as multimeric molecular complexes involved in BCR-activated calcium entry. Using coprecipitation of native CD20 with tagged or truncated forms of the molecule, we provide here direct evidence of CD20 homo-oligomerization into tetramers. Additionally, the function of CD20 was explored by examining its association with surface-labeled and intracellular proteins before and after BCR signaling. Two major surface-labeled proteins that coprecipitated with CD20 were identified as the heavy and light chains of cell surface IgM, the antigen-binding components of the BCR. After activation, BCR-CD20 complexes dissociated, and phosphoproteins and calmodulin-binding proteins were transiently recruited to CD20. These data provide new evidence of the involvement of CD20 in signaling downstream of the BCR and, together with the previously described involvement of CD20 in calcium influx, the first evidence of physical coupling of the BCR to a calcium entry pathway.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens, CD20 / immunology*
  • Antigens, CD20 / metabolism
  • B-Lymphocytes / immunology*
  • B-Lymphocytes / metabolism
  • Calcium / immunology
  • Calcium / metabolism
  • Calcium Signaling / physiology*
  • Calmodulin-Binding Proteins / immunology*
  • Calmodulin-Binding Proteins / metabolism
  • Cell Line, Tumor
  • Cell Membrane / immunology
  • Cell Membrane / metabolism
  • Immunoglobulin M / immunology
  • Immunoglobulin M / metabolism
  • Lymphocyte Activation / physiology*
  • Mice
  • Protein Structure, Quaternary / physiology
  • Receptors, Antigen, B-Cell / immunology*
  • Receptors, Antigen, B-Cell / metabolism

Substances

  • Antigens, CD20
  • Calmodulin-Binding Proteins
  • Immunoglobulin M
  • Receptors, Antigen, B-Cell
  • Calcium