Folding and function of I domain-deleted Mac-1 and lymphocyte function-associated antigen-1

J Biol Chem. 2000 Jul 21;275(29):21877-82. doi: 10.1074/jbc.M908868199.

Abstract

In those integrins that contain it, the I domain is a major ligand recognition site. The I domain is inserted between beta-sheets 2 and 3 of the predicted beta-propeller domain of the integrin alpha subunit. We deleted the I domain from the integrin alpha(M) and alpha(L) subunits to give I-less Mac-1 and lymphocyte function-associated antigen-1 (LFA-1), respectively. The I-less alpha(M) and alpha(L) subunits were expressed in association with the wild-type beta(2) subunit on the surface of transfected cells and bound to all the monoclonal antibodies mapped to the putative beta-propeller and C-terminal regions of the alpha(M) and alpha(L) subunits, suggesting that the folding of these domains is independent of the I domain. I-less Mac-1 bound to the ligands iC3b and factor X, but this binding was reduced compared with wild-type Mac-1. In contrast, I-less Mac-1 did not bind to fibrinogen or denatured bovine serum albumin. Binding to iC3b and factor X by I-less Mac-1 was inhibited by the function-blocking antibody CBRM1/32, which binds to the beta-propeller domain of the alpha(M) subunit. I-less LFA-1 did not bind its ligands intercellular adhesion molecule-1 and -3. Thus, the I domain is not essential for the folding, heterodimer formation, and surface expression of Mac-1 and LFA-1 and is required for binding to some ligands, but not others.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Binding Sites
  • Cattle
  • Cell Line
  • Humans
  • Ligands
  • Lymphocyte Function-Associated Antigen-1 / chemistry*
  • Lymphocyte Function-Associated Antigen-1 / genetics*
  • Lymphocyte Function-Associated Antigen-1 / metabolism
  • Macrophage-1 Antigen / chemistry*
  • Macrophage-1 Antigen / genetics*
  • Macrophage-1 Antigen / metabolism
  • Protein Folding*
  • Sequence Deletion
  • Structure-Activity Relationship

Substances

  • Ligands
  • Lymphocyte Function-Associated Antigen-1
  • Macrophage-1 Antigen