Calcium-triggered selective intermembrane exchange of phospholipids by the lung surfactant protein SP-A.
It is shown that human lung surfactant protein (SP-A) mediates selective exchange of phospholipid probes with unlabeled phospholipid in excess vesicles in the presence of calcium and NaCl. The exchange occurs without leakage of vesicle contents, or transbilayer movement (flip-flop) of the phospholipid probes, or fusion of vesicles. Individual steps preceding the exchange are dissected by a combination of protocols, and the results are operationally interpreted in terms of a model where a calcium-dependent change in SP-A triggers aggregation of vesicles followed by probe exchange between the vesicles in contact through SP-A. The contacts remain stable in the presence of calcium; i.e., the vesicles in contact do not change their partners on the time scale of several minutes. The binding of SP-A to vesicles and the aggregation of vesicles are rapid, and the aggregation is rapidly reversed by EGTA; i.e., both the forward and reverse aggregation reactions are complete in about 1 min. The exchange rate of the various probes between aggregated vesicles below 1 mM calcium in the presence of NaCl shows selectivity, i.e., a modest dependence on the net anionic charge on vesicles and for the headgroup of the probe. Exchange with lower selectivity is seen at >2 mM Ca in the absence of NaCl. SP-A binding to vesicles does not show an absolute specificity for the phospholipid structure, but the time course of the subsequent changes does. The results suggest that SP-A contacts between phospholipid interfaces could mediate the exchange of phospholipid species (trafficking and sorting) between lung surfactant pools in the hypophase and all accessible phospholipid interfaces of the alveolar space.
Department of Chemistry and Biochemistry, University of Delaware, Newark 19716, USA.
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Y
Cajal
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Dodia
A B
AB
Fisher
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MK
Jain
D002118:Calcium / Q000502:physiology*; D002122:Calcium Chloride / Q000494:pharmacology; D002462:Cell Membrane / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D005454:Fluorescence Polarization; D005456:Fluorescent Dyes / Q000378:metabolism; D006801:Humans; D008081:Liposomes / Q000378:metabolism; D008168:Lung / Q000378:metabolism; D008563:Membrane Lipids / Q000737:chemistry / Q000378:metabolism*; D010713:Phosphatidylcholines / Q000378:metabolism; D010714:Phosphatidylethanolamines / Q000378:metabolism; D011510:Proteolipids / Q000378:metabolism* / Q000494:pharmacology; D037662:Pulmonary Surfactant-Associated Protein A; D037661:Pulmonary Surfactant-Associated Proteins; D011663:Pulmonary Surfactants / Q000378:metabolism* / Q000494:pharmacology; D011721:Pyrenes / Q000378:metabolism; D013109:Sphingomyelins / Q000378:metabolism; D014364:Tryptophan / Q000378:metabolism; D014867:Water
D005456:Fluorescent Dyes; D008081:Liposomes; D008563:Membrane Lipids; C053019:N-(10-(1-pyrenyl)decanoyl)sphingomyelin; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; D011510:Proteolipids; D037662:Pulmonary Surfactant-Associated Protein A; D037661:Pulmonary Surfactant-Associated Proteins; D011663:Pulmonary Surfactants; D011721:Pyrenes; D013109:Sphingomyelins; D014867:Water; C042077:N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)phosphatidylethanolamine; C021248:1-palmitoyl-2-pyrenedecanoylphosphatidylcholine; D014364:Tryptophan; D002122:Calcium Chloride; D002118:Calcium