Abstract
The solubility of hydrophobic pulmonary surfactant proteins in different organic solvents and organic solvent/water combinations has been analyzed. Three organic solvents have been selected: methanol (MetOH), acetonitrile (ACN) and trifluorethanol (TFE). Porcine SP-B showed very similar calculated secondary structure when dissolved in methanol, 60% ACN or 70% TFE and reconstituted in lysophosphatidylcholine (LPC) micelles or dipalmitoylphosphatidylcholine (DPPC) vesicles, as deduced from circular dichroism studies. SP-B was calculated to posses around 45% of α-helix in all these systems. The fluorescence emission spectrum of SP-B has been also characterized in aqueous solvents and lipids. It always showed a splitting of the tryptophan contribution into two components with different emission maxima. SP-C had a very different structure in 80% ACN or 70% TFE. While α-helix was the main secondary structure of SP-C in ACN/water mixtures — around 50% -, it had almost exclusively β-structure when dissolved in 70% TFE. The CD spectrum of SP-C in TFE showed dependence on the protein concentration, suggesting that protein-protein interactions could be important in this β-conformation. SP-C reconstituted in LPC micelles or DPPC vesicles had a CD spectrum qualitatively similar to that one in aqueous ACN, with a dominant α-helical structure. The α-helical content of SP-C in micelles of LPC and vesicles of DPPC, 60 and 70%, respectively, was calculated to be higher than the α-helical content of the protein dissolved in any aqueous organic solvent.
Published Version
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