Specific binding of human C-reactive protein towards supported monolayers of binary and engineered phospholipids.

Circulating C-reactive protein (CRP) recognizes altered plasma membranes and activates complements systems in the acute phase of inflammation and infection in human. We have shown previously the calcium-independent adsorption of CRP toward 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) and lysophosphatidylcholine (LPC) on supported phospholipid monolayers. Here, we extended our study to other phospholipids and additives to elucidate the pattern recognition of CRP using a surface plasmon resonance biosensor. Surface density and lateral fluidity depended on the type of phospholipids in the monolayers as characterized by SPR and fluorescence recovery after photobleaching measurements. CRP recognized 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-l-serine (POPS) and 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-(1'-rac-glycerol) (POPG) in the supported POPC monolayers without calcium at pH 7.4 and 5.5. As opposed to LPC, CRP did not recognize 3-sn-lysophosphatidylethanolamine in the POPC monolayers in calcium-free conditions. While, the addition of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE) or sphingomyelin to supported POPC monolayers blocked CRP adsorption. Calcium-dependent CRP binding was observed only at pH 5.5 on supported monolayers of engineered phospholipids with inverted headgroups relative to POPC. The complement 1q (C1q) protein recognized the active form of CRP on the supported phospholipid monolayers. The discovery of CRP recognition with these phospholipids aids our understanding of the activation dynamics of CRP with phospholipid-based biomaterials when used during the acute phase.