title: Synthesis and structural characterization of human-identical lung surfactant SP-C protein.
abstract: An efficient synthesis for human-identical lung surfactant protein SP-C is described with a semi-automated solid phase synthesizer using Fmoc chemistry. Double coupling and acetic anhydride capping procedures were employed for synthetic cycles within the highly hydrophobic C-terminal domain of SP-C. Isolation of the protein was performed by mild cleavage and deprotection conditions and subsequent HPLC purification yielding a highly homogeneous protein as established by sequence determination, electrospray, plasma desorption and MALDI mass spectrometry. A general method has been employed for the preparation of Cys-palmitoylated protein by using temporary Cys(tButhio) protection, in situ deprotection with beta-mercaptoethanol and selective palmitoylation of resin-bound SP-C. The mild synthesis and isolation conditions provide SP-C with a high alpha-helical content, comparable to that of the natural SP-C, as assessed by CD spectra. Furthermore, first biophysical data indicate a surfactant activity comparable to that of the natural protein.
authors.affiliation: Fakultät für Chemie, Universität Konstanz, Germany.
authors.forename: P
authors.initials: P
authors.lastname: Mayer-Fligge
authors.forename: J
authors.initials: J
authors.lastname: Volz
authors.forename: U
authors.initials: U
authors.lastname: Krüger
authors.forename: E
authors.initials: E
authors.lastname: Sturm
authors.forename: W
authors.initials: W
authors.lastname: Gernandt
authors.forename: K P
authors.initials: KP
authors.lastname: Schäfer
authors.forename: M
authors.initials: M
authors.lastname: Przybylski
mesh_terms: D000595:Amino Acid Sequence; D002851:Chromatography, High Pressure Liquid; D002942:Circular Dichroism; D006801:Humans; D008969:Molecular Sequence Data; D011510:Proteolipids / Q000138:chemical synthesis* / Q000737:chemistry*; D011663:Pulmonary Surfactants / Q000138:chemical synthesis* / Q000737:chemistry*
chemical_list: D011510:Proteolipids; D011663:Pulmonary Surfactants