title: Pulmonary surfactant protein-A (SP-A) restores the surface properties of surfactant after oxidation by a mechanism that requires the Cys6 interchain disulfide bond and the phospholipid binding domain.
abstract: Reactive oxygen species produced by activated leuko-cytes in the alveolar epithelial lining fluid have been implicated in the inactivation of pulmonary surfactant and the impairment of lung function. Oxidation of bovine lipid extract surfactant (BLES), a therapeutic surfactant, with hypochlorous acid (H-BLES) or the Fenton reaction (F-BLES) led to temporary increases in conjugated dienes and formation of malondialdehyde and 4-hydroxy-2-nonenal. Electrospray ionization mass spectrometry revealed the appearance of lipid hydroperoxides, peroxides, lysophospholipids, and free fatty acids. Captive bubble tensiometer studies of H-BLES demonstrated prolonged adsorption times, film instability at low surface tensions during film compression, and reduced respreadability during film expansion. F-BLES exhibited prolonged adsorption times, a marked effect on increasing compressibility during compression, and a lesser effect on reducing respreadability on expansion. Addition of native bovine or rat surfactant-associated protein A (SP-A) reversed the effects of oxidation on surfactant biophysical properties. Studies using mutant recombinant rat SP-As indicated that an intact carbohydrate recognition domain and disulfide-dependent oligomeric assembly are critical for these effects, but the collagen-like region is not required. We conclude that SP-A can reverse the detrimental effects of surfactant oxidation on the biophysical properties of surfactant, by a mechanism that is dependent on interchain disulfide bond formation and the C-terminal domains of the protein.
authors.affiliation: Departments of Obstetrics/Gynecology and Biochemistry, Canadian Institutes of Health Research Group in Fetal and Neonatal Health and Development, University of Western Ontario, London N6A 5A5, Canada.
authors.forename: Karina
authors.initials: K
authors.lastname: Rodriguez Capote
authors.forename: Francis X
authors.initials: FX
authors.lastname: McCormack
authors.forename: Fred
authors.initials: F
authors.lastname: Possmayer
mesh_terms: D000818:Animals; D001665:Binding Sites / Q000502:physiology; D002417:Cattle; D003545:Cysteine / Q000378:metabolism; D004220:Disulfides / Q000378:metabolism; D005609:Free Radicals / Q000378:metabolism; D006861:Hydrogen Peroxide; D006997:Hypochlorous Acid; D007501:Iron; D010084:Oxidation-Reduction; D010743:Phospholipids / Q000378:metabolism*; D037662:Pulmonary Surfactant-Associated Protein A / Q000737:chemistry* / Q000378:metabolism*; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism; D021241:Spectrometry, Mass, Electrospray Ionization; D013501:Surface-Active Agents / Q000378:metabolism
chemical_list: D004220:Disulfides; C045076:Fenton's reagent; D005609:Free Radicals; D010743:Phospholipids; D037662:Pulmonary Surfactant-Associated Protein A; D011994:Recombinant Proteins; D013501:Surface-Active Agents; D006997:Hypochlorous Acid; D006861:Hydrogen Peroxide; D007501:Iron; D003545:Cysteine