Synthesis, purification and surface activities of the human pulmonary surfactant protein-C (SP-C) analogue, SP-CL16 (6-28).
We previously reported that a human analogue of pulmonary surfactant protein-C (SP-C), SP-CL16 (6-28), with 23 residues (Fig. 1) was the most active analogue in a reconstituted lipid mixture and had the shortest chain among the poly-leucine-analogues examined. In the present study, we examined a new method of preparing this analogue, that is, stepwise solid-phase synthesis employing the Fmoc method followed by centrifugal partition chromatography (CPC) using an n-hexane/CH3OH/H2O/trifluoroacetic acid (TFA) (1000: 1000:1:2, v/v) solvent system according to the descending method. The synthetic peptides were identified by matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry in search of activity to improve the in vitro surface activity of a ternary lipid mixture composed of dipalmitoylphosphatidylcholine, egg-phosphatidylglycerol and palmitic acid (75:25:10, w/w) in a Langmuir-Wilhelmy surface balance. SP-CL16 (6-28) seemed comparable in surface activity with Surfacten (Surfactant-TA), a modified surfactant preparation which has been used for the treatment of respiratory distress syndrome.
Otsubo
E
E
Research Center, Mitsubishi-Tokyo Pharmaceuticals, Inc, Kisarazu, Chiba, Japan. 4600423@mitsubishi-pharm.co.jp
Takei
T
T
Nomura
M
M
D000595:Amino Acid Sequence; D006801:Humans; D008969:Molecular Sequence Data; D011510:Proteolipids / Q000138:chemical synthesis* / Q000737:chemistry / Q000302:isolation & purification*; D011663:Pulmonary Surfactants / Q000138:chemical synthesis* / Q000737:chemistry / Q000302:isolation & purification*; D013499:Surface Properties; D013501:Surface-Active Agents / Q000138:chemical synthesis* / Q000737:chemistry / Q000302:isolation & purification*
D011510:Proteolipids; D011663:Pulmonary Surfactants; D013501:Surface-Active Agents