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Volumn 273, Issue 5279, 1996, Pages 1196-1199

Phase and morphology changes in lipid monolayers induced by SP-B protein and its amino-terminal peptide

Author keywords

[No Author keywords available]

Indexed keywords

DIPALMITOYLPHOSPHATIDYLCHOLINE; LUNG SURFACTANT; PALMITIC ACID; PHOSPHATIDYLCHOLINE; PHOSPHATIDYLGLYCEROL; SURFACTANT TA; SYNTHETIC PEPTIDE;

EID: 0029789206     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.273.5279.1196     Document Type: Article
Times cited : (168)

References (47)
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    • 1-25 were synthesized and analyzed as described in (10), except that, for the human 78-residue protein SP-B, we used an ABl 431A peptide synthesizer (Applied Biosystem, Foster City, CA) applying FastMoc chemistry (11). Amino acid residues 26 to 59 of SP-B were double-coupled; all other residues were single-coupled. After cleavage from the resin, we purified the peptide by reverse-phase highperformance liquid chromatography (HPLC) with a Vydac C4 column (Vydac, Hesperia, CA), using a water-acetonitrile gradient containing 0.1% trifluoroacetic acid. The formation of disulfide bonds was facilitated with the use of EKATHIOX resin (Ekagen, Palo Alto, CA). Oxidatbn with EKATHIOX resin was carried out by addition of a 1 mM peptide solution of trifluoroethanol:water (8:2 v/v) to a 10-fold molar excess of the resin-active group to peptide thiol. The reaction was allowed to proceed for 6 hours before the peptide in solution was separated from the resin by centrifugation (1000g, 10 min). The monomeric oxidized state was confirmed by matrix-assisted laser desorptiontime of flight (MALDI-TOF) mass spectroscopy. The conformation of SP-B in the PA multilayer was largely helical as determined by Fourier-transform infrared spectroscopy measurements.
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    • We thank D. K. Schwartz, C. Knobler, R. Stuber, and A. Weinberg for help with trough construction; K. Faull and R. Stevens for help with the mass spectral analysis; and L. Gordon for his reading of the manuscript. The work of J.A.Z., M.M.L., and K.Y.C.L. was supported by NIH grant HL51177-02; K.Y.C.L. received a University of California President's postdoctoral fellowship; and M.M.L. received a University of California Regents graduate fellowship. The work of A.J.W. was supported by NIH grant HL55534 and NIH Small Equipment grant GM 50483.


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