title: Use of human surfactant low molecular weight apoproteins in the reconstitution of surfactant biologic activity.
abstract: Two low molecular weight (LMW) apoproteins were isolated from human pulmonary surfactant. SDS polyacrylamide gel analysis showed one protein (SP 18) to have an apparent molecular weight of 18,000 when unreduced and 9,000 D after reduction. The second protein (SP 9) migrated at approximately 9,000 D in the presence or absence of reducing agents. Both proteins contain a high number of hydrophobic amino acids. The NH2-terminal sequence of SP 18 was determined to be: NH2-phe-pro-ile-pro-leu-pro-tyr-. A cDNA clone isolated from a human adult lung cDNA library contained a long open reading frame encoding at an internal position the human SP 18 amino-terminal sequence. Mixtures of phospholipids (PL) and SP 9 and SP 18 were assessed for their capacity to reduce surface tensions on a pulsating bubble surfactometer. The addition of 1% apoprotein resulted in a reduction of surface tension after 15 s from 42.9 dyn/cm for PL alone to 16.7 and 6.3 dyn/cm for preparations containing SP 9 and SP 18, respectively. In vivo assessment of reconstituted surfactant activity was performed in fetal rabbits. Reconstituted surfactant consisting of PL + 0.5% SP 18 instilled intratracheally at delivery resulted in a marked increase in lung compliance, while the incorporation of 0.5% SP 9 yielded a moderate increase. These data show the ability to produce biologically active surfactant by the addition of isolated LMW apoproteins to defined PL.
authors.affiliation: Department of Immunology, Research Institute of Scripps Clinic, La Jolla, California 92037.
authors.forename: S D
authors.initials: SD
authors.lastname: Revak
authors.forename: T A
authors.initials: TA
authors.lastname: Merritt
authors.forename: E
authors.initials: E
authors.lastname: Degryse
authors.forename: L
authors.initials: L
authors.lastname: Stefani
authors.forename: M
authors.initials: M
authors.lastname: Courtney
authors.forename: M
authors.initials: M
authors.lastname: Hallman
authors.forename: C G
authors.initials: CG
authors.lastname: Cochrane
mesh_terms: D000595:Amino Acid Sequence; D000653:Amniotic Fluid / Q000032:analysis; D000818:Animals; D001059:Apoproteins / Q000302:isolation & purification* / Q000494:pharmacology; D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA / Q000302:isolation & purification; D006801:Humans; D008170:Lung Compliance / Q000187:drug effects; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011650:Pulmonary Alveoli / Q000503:physiopathology; D037661:Pulmonary Surfactant-Associated Proteins; D011663:Pulmonary Surfactants / Q000302:isolation & purification* / Q000494:pharmacology; D011817:Rabbits
chemical_list: D001059:Apoproteins; D037661:Pulmonary Surfactant-Associated Proteins; D011663:Pulmonary Surfactants; C046616:pulmonary surfactant apoprotein; D004247:DNA