Deacylated pulmonary surfactant protein SP-C transforms from alpha-helical to amyloid fibril structure via a pH-dependent mechanism: an infrared structural investigation.
Bovine pulmonary surfactant protein C (SP-C) is a hydrophobic, alpha-helical membrane-associated lipoprotein in which cysteines C4 and C5 are acylated with palmitoyl chains. Recently, it has been found that the alpha-helix form of SP-C is metastable, and under certain circumstances may transform from an alpha-helix to a beta-strand conformation that resembles amyloid fibrils. This transformation is accelerated when the protein is in its deacylated form (dSP-C). We have used infrared spectroscopy to study the structure of dSP-C in solution and at membrane interfaces. Our results show that dSP-C transforms from an alpha-helical to a beta-type amyloid fibril structure via a pH-dependent mechanism. In solution at low pH, dSP-C is alpha-helical in nature, but converts to an amyloid fibril structure composed of short beta-strands or beta-hairpins at neutral pH. The alpha-helix structure of dSP-C is fully recoverable from the amyloid beta-structure when the pH is once again lowered. Attenuated total reflectance infrared spectroscopy of lipid-protein monomolecular films showed that the fibril beta-form of dSP-C is not surface-associated at the air-water interface. In addition, the lipid-associated alpha-helix form of dSP-C is only retained at the surface at low surface pressures and dissociates from the membrane at higher surface pressures. In situ polarization modulation infrared spectroscopy of protein and lipid-protein monolayers at the air-water interface confirmed that the residual dSP-C helix conformation observed in the attenuated total reflectance infrared spectra of transferred films is randomly or isotropically oriented before exclusion from the membrane interface. This work identifies pH as one of the mechanistic causes of amyloid fibril formation for dSP-C, and a possible contributor to the pathogenesis of pulmonary alveolar proteinosis.
D015060:1,2-Dipalmitoylphosphatidylcholine / Q000737:chemistry; D000215:Acylation; D000682:Amyloid / Q000737:chemistry*; D000818:Animals; D002417:Cattle; D006863:Hydrogen-Ion Concentration; D008051:Lipid Bilayers / Q000737:chemistry*; D046911:Macromolecular Substances; D008563:Membrane Lipids / Q000737:chemistry*; D008970:Molecular Weight; D010715:Phosphatidylglycerols / Q000737:chemistry; D011487:Protein Conformation; D020033:Protein Isoforms / Q000737:chemistry; D017433:Protein Structure, Secondary; D037721:Pulmonary Surfactant-Associated Protein C / Q000737:chemistry*; D012996:Solutions; D013055:Spectrophotometry, Infrared
D000682:Amyloid; D008051:Lipid Bilayers; D046911:Macromolecular Substances; D008563:Membrane Lipids; D010715:Phosphatidylglycerols; D020033:Protein Isoforms; D037721:Pulmonary Surfactant-Associated Protein C; D012996:Solutions; D015060:1,2-Dipalmitoylphosphatidylcholine; C030345:1,2-dipalmitoylphosphatidylglycerol