Modification of tryptophan and methionine residues is implicated in the oxidative inactivation of surfactant protein B.
Exposing BLES (bovine lipid extract surfactant), a clinical surfactant, to reactive oxygen species (ROS) alters surfactant protein B (SP-B), as indicated by Coomassie Blue staining, silver staining, and Western analysis. Hypochlorous acid (HOCl) treatment leads to elevated maximum surface tension (gammamax) and a deterioration in minimum gamma (gammamin) during surface area cycling. Fenton reaction resulted in immediate increases in gammamin and gammamax. Intrinsic fluorescence measurements indicated Fenton, but not HOCl, induced conversion of Trp9 of SP-B to hydroxyTrp (OHTrp), N-formylkynurenine (NFKyn), and kynurenine (Kyn). Electrospray ionization mass spectrometry (ESI-MS) revealed molecular weight alterations consistent with oxidation of Met (HOCl, Fenton) and Trp (Fenton) residues. Oxidative alterations to Met29 and Met65 (HOCl, Fenton) and to Trp9 (OHTrp with HOCL and NFKyn plus Kyn with Fenton) were confirmed by matrix-assisted laser desorption mass spectrometry (MALDI-MS) studies on SP-B tryptic fragments. Some Met oxidation was observed with control SP-B. When taken together with captive bubble tensiometer measurements, these studies suggest that Met oxidation of SP-B by HOCl or Fenton interferes with phospholipid respreading during compression-expansion of surfactant films, while Fenton oxidation, which produces more extensive Met oxidation and disruption of the indole ring of Trp9, further abrogated the ability of such films to attain low surface tensions during compression. These studies provide insight into the manner by which ROS generated during acute lung injury and the acute respiratory distress syndrome act to inhibit not only endogenous surfactant but also therapeutic surfactants administered to counteract these conditions.
Manzanares
Dahis
D
Department of Obstetrics and Gynaecology, Schulich School of Medicine and Dentistry, The University of Western Ontario, London, ON, Canada N6A 5C1.
Rodriguez-Capote
Karina
K
Liu
Suya
S
Haines
Thomas
T
Ramos
Yudith
Y
Zhao
Lin
L
Doherty-Kirby
Amanda
A
Lajoie
Gilles
G
Possmayer
Fred
F
D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D006861:Hydrogen Peroxide / Q000737:chemistry; D006997:Hypochlorous Acid / Q000737:chemistry; D007501:Iron / Q000737:chemistry; D008715:Methionine / Q000737:chemistry / Q000378:metabolism*; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D010743:Phospholipids / Q000737:chemistry; D037701:Pulmonary Surfactant-Associated Protein B / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism*; D017382:Reactive Oxygen Species / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014364:Tryptophan / Q000737:chemistry / Q000378:metabolism*
C045076:Fenton's reagent; D010743:Phospholipids; D037701:Pulmonary Surfactant-Associated Protein B; D017382:Reactive Oxygen Species; D006997:Hypochlorous Acid; D014364:Tryptophan; D008715:Methionine; D006861:Hydrogen Peroxide; D007501:Iron