|Title||Use of single point mutations in domain I of beta 2-glycoprotein I to determine fine antigenic specificity of antiphospholipid autoantibodies|
|Publication Type||Journal Article|
|Year of Publication||2002|
|Authors||Iverson, GM, Reddel, S, Victoria, EJ, Cockerill, KA, Wang, YX, Marti-Renom, MA, Sali, A, Marquis, DM, Krilis, SA, Linnik, MD|
|Keywords||Amino Acid Substitution/genetics Antibodies; Antibody/genetics Binding; Antiphospholipid/blood/*metabolism Antibodies; Competitive/genetics/immunology Enzyme-Linked Immunosorbent Assay/methods Epitopes/analysis/*immunology/metabolism Glycine/genetics Glycoproteins/biosynthesis/*genetics/*immunology/isolation & purification/metabolism Humans Models; Molecular Peptide Fragments/genetics/immunology/isolation & purification/metabolism *Point Mutation Protein Structure; Monoclonal/blood/metabolism Antiphospholipid Syndrome/immunology Arginine/genetics *Binding Sites; Tertiary/genetics Recombinant Proteins/biosynthesis/immunology/isolation & purification/metabolism Static Electricity beta 2-Glycoprotein I|
Autoantibodies against beta(2)-glycoprotein I (beta(2)GPI) appear to be a critical feature of the antiphospholipid syndrome (APS). As determined using domain deletion mutants, human autoantibodies bind to the first of five domains present in beta(2)GPI. In this study the fine detail of the domain I epitope has been examined using 10 selected mutants of whole beta(2)GPI containing single point mutations in the first domain. The binding to beta(2)GPI was significantly affected by a number of single point mutations in domain I, particularly by mutations in the region of aa 40-43. Molecular modeling predicted these mutations to affect the surface shape and electrostatic charge of a facet of domain I. Mutation K19E also had an effect, albeit one less severe and involving fewer patients. Similar results were obtained in two different laboratories using affinity-purified anti-beta(2)GPI in a competitive inhibition ELISA and with whole serum in a direct binding ELISA. This study confirms that anti-beta(2)GPI autoantibodies bind to domain I, and that the charged surface patch defined by residues 40-43 contributes to a dominant target epitope.
Iverson, G Michael Reddel, Stephen Victoria, Edward J Cockerill, Keith A Wang, Ying-Xia Marti-Renom, Marc A Sali, Andrej Marquis, David M Krilis, Steven A Linnik, Matthew D GM54762/GM/NIGMS NIH HHS/United States Research Support, Non-U.S. Gov’t Research Support, U.S. Gov’t, P.H.S. United States Journal of immunology (Baltimore, Md. : 1950) J Immunol. 2002 Dec 15;169(12):7097-103.