title: Topology and lipid selectivity of pulmonary surfactant protein SP-B in membranes: Answers from fluorescence.
abstract: Contradictory results have been reported with respect to the depth of penetration and the orientation of pulmonary surfactant protein SP-B in phospholipid membranes and its relative selectivity to interact with anionic over zwitterionic phospholipid species. In the present study we have re-evaluated lipid-protein interactions of SP-B by analysing Forster resonance energy transfer (FRET) efficiencies, obtained from time-resolved measurements, from the single tryptophan in SP-B to different fluorescently labelled phospholipids in matrix bilayers made of either pure phosphatidylcholine (POPC) or the full lipid extract obtained from purified surfactant. In the background of POPC membranes SP-B exhibits a certain level of selectivity for anionic fluorescent phospholipids over the corresponding zwitterionic analogues, but apparently no preference for phosphatidylglycerol over other anionic species such as phosphatidylserine. No selectivity was detected in membranes made of full surfactant lipids, indicating that specific lipid-protein binding sites could already be occupied by endogenous anionic phospholipids. Furthermore, we have analysed the fit of two different models of how SP-B could be orientated with respect to phospholipid membrane surfaces to the FRET data. The FRET results are consistent with topology models in which the protein has a superficial orientation, with no regions of exclusion by the protein to the access of phospholipids, both in POPC membranes and in membranes made of the whole surfactant lipid fraction. This discards a deep penetration of the protein into the core of bilayers and suggests that most hydrophobic segments of SP-B could participate in protein-protein instead of lipid-protein interactions.
authors.affiliation: Dept. Bioquimica, Fac. Biologia, Universidad Complutense, 28040 Madrid, Spain.
authors.forename: Elisa J
authors.initials: EJ
authors.lastname: Cabré
authors.forename: Luís M S
authors.initials: LM
authors.lastname: Loura
authors.forename: Alexander
authors.initials: A
authors.lastname: Fedorov
authors.forename: Jesus
authors.initials: J
authors.lastname: Perez-Gil
authors.forename: Manuel
authors.initials: M
authors.lastname: Prieto
mesh_terms: D009327:4-Chloro-7-nitrobenzofurazan / Q000737:chemistry; D000465:Algorithms; D000818:Animals; D001665:Binding Sites; D001667:Binding, Competitive; D031541:Fluorescence Resonance Energy Transfer; D020404:Glycerophospholipids / Q000737:chemistry* / Q000378:metabolism; D008051:Lipid Bilayers / Q000737:chemistry* / Q000378:metabolism; D008055:Lipids / Q000737:chemistry*; D008958:Models, Molecular; D010713:Phosphatidylcholines / Q000737:chemistry / Q000378:metabolism; D010715:Phosphatidylglycerols / Q000737:chemistry / Q000378:metabolism; D010718:Phosphatidylserines / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D037701:Pulmonary Surfactant-Associated Protein B / Q000737:chemistry* / Q000378:metabolism; D013552:Swine
chemical_list: D020404:Glycerophospholipids; D008051:Lipid Bilayers; D008055:Lipids; D010713:Phosphatidylcholines; D010715:Phosphatidylglycerols; D010718:Phosphatidylserines; D037701:Pulmonary Surfactant-Associated Protein B; D009327:4-Chloro-7-nitrobenzofurazan