Activity-regulating structural changes and autoantibody epitopes in transglutaminase 2 assessed by hydrogen/deuterium exchange

Proc Natl Acad Sci U S A. 2014 Dec 2;111(48):17146-51. doi: 10.1073/pnas.1407457111. Epub 2014 Nov 17.

Abstract

The multifunctional enzyme transglutaminase 2 (TG2) is the target of autoantibodies in the gluten-sensitive enteropathy celiac disease. In addition, the enzyme is responsible for deamidation of gluten peptides, which are subsequently targeted by T cells. To understand the regulation of TG2 activity and the enzyme's role as an autoantigen in celiac disease, we have addressed structural properties of TG2 in solution by using hydrogen/deuterium exchange monitored by mass spectrometry. We demonstrate that Ca(2+) binding, which is necessary for TG2 activity, induces structural changes in the catalytic core domain of the enzyme. Cysteine oxidation was found to abolish these changes, suggesting a mechanism whereby disulfide bond formation inactivates the enzyme. Further, by using TG2-specific human monoclonal antibodies generated from intestinal plasma cells of celiac disease patients, we observed that binding of TG2 by autoantibodies can induce structural changes that could be relevant for the pathogenesis. Detailed mapping of two of the main epitopes targeted by celiac disease autoantibodies revealed that they are located adjacent to each other in the N-terminal part of the TG2 molecule.

Keywords: autoantibodies; celiac disease; hydrogen/deuterium exchange; transglutaminase 2.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Autoantibodies / immunology*
  • Autoantibodies / metabolism
  • Binding Sites / genetics
  • Calcium / chemistry
  • Calcium / metabolism
  • Celiac Disease / immunology
  • Celiac Disease / metabolism
  • Celiac Disease / pathology
  • Deuterium Exchange Measurement / methods*
  • Disulfides / chemistry
  • Disulfides / metabolism
  • Electrophoresis, Polyacrylamide Gel
  • Epitope Mapping / methods
  • Epitopes / chemistry
  • Epitopes / immunology*
  • Epitopes / metabolism
  • GTP-Binding Proteins / chemistry
  • GTP-Binding Proteins / genetics
  • GTP-Binding Proteins / immunology*
  • Humans
  • Intestinal Mucosa / metabolism
  • Intestines / immunology
  • Intestines / pathology
  • Mass Spectrometry / methods
  • Models, Molecular
  • Molecular Sequence Data
  • Mutation
  • Plasma Cells / immunology
  • Plasma Cells / metabolism
  • Protein Binding
  • Protein Glutamine gamma Glutamyltransferase 2
  • Protein Structure, Tertiary
  • Transglutaminases / chemistry
  • Transglutaminases / genetics
  • Transglutaminases / immunology*

Substances

  • Autoantibodies
  • Disulfides
  • Epitopes
  • Protein Glutamine gamma Glutamyltransferase 2
  • Transglutaminases
  • GTP-Binding Proteins
  • Calcium