Calcium-independent binding of human C-reactive protein to lysophosphatidylcholine in supported planar phospholipid monolayers

摘要

Details describing the molecular dynamics of inflammation biomarker human C-reactive protein (CRP) on plasma membranes containing bioactive lipid lysophosphatidylcholine (LPC) remain elusive. Here, we measured the binding kinetics of CRP to supported phospholipid monolayers deposited on an alkanethiol self -assembled monolayer on a planar gold substrate using surface plasmon resonance. Surprisingly, CRP binding to supported 1-palmitoy1-2-oleoyl-sn-glycero-3-phosphocholine (POPC)/LPC monolayers was calcium-independent although CRP binding to supported POPC monolayers was calcium-dependent. Binding inhibition assays indicate a specific interaction between CRP and the glycerophosphate group in LPC in the absence of calcium ions. Binding experiments on supported POPO -palmitoy1-2-oleoylsn-glycero-3-phospho-(1'-rac-glycerol) monolayers further validated calcium-independent binding of CRP through the glycerophosphate moiety. Docking analysis predicted a new binding site for LPC in the absence of calcium ions, which is located on the opposite side of the known binding site for PC of cyclic pentameric CRP. These results using model plasma membranes should aid our understanding of the activation dynamics of CRP in altered local microenvironments of inflammation and infection.