Evolutionary conservation of the lipopolysaccharide binding site of β2-glycoprotein I

Abstract

Summaryβ2-Glycoprotein I (β2GPI) is a highly abundant plasma protein and the major antigen for autoantibodies in the antiphospholipid syndrome. Recently, we have described a novel function of β2GPI as scavenger of lipopolysaccharide (LPS). With this in mind we investigated the conservation of β2GPI in vertebrates and set out to identify the binding site of LPS within β2GPI. The genome sequences of 42 species were surveyed. Surface plasmon resonance (SPR) was performed with peptides to characterise the binding site of β2GPI for LPS. β2GPI could be identified in most tested vertebrates with a high overall amino acid homology of 80% or more in mammals. SPR revealed that a synthesised peptide (LAFWKTDA) from domain V of β2GPI was able to compete for binding of β2GPI to LPS. The AFWKTDA sequence was completely conserved in all mammals. The peptide containing the LPS binding site attenuated the inhibition by β2GPI in a cellular model of LPS-induced tissue factor expression. Other important sites, such as the binding site for anionic phospholipids and the antiphospholipid antibody binding epitope, were also preserved. β2GPI is highly conserved across the animal kingdom, which suggests that the function of β2GPI may be more important than anticipated.