Separation of subfractions of the hydrophobic components of calf lung surfactant

Stephen (Steve) Hall, Z. Wang, R. H. Notter

Research output: Contribution to journalArticle

60 Citations (Scopus)

Abstract

This study reports the biochemical separation of the hydrophobic constituents of calf lung surfactant into separate fractions from which specific components are excluded. Gel permeation chromatography on LH-20 with acidified chloroform-methanol separated the constituents of calf lung surfactant extract (CLSE) into fractions containing purified phospholipids (PPL), the neutral lipids and phospholipids (N and PL), or the hydrophobic surfactant proteins (SP)-B and -C together with the phospholipids (SP and PL). Extraction of acid to prevent phospholipid degradation after separation reduced recovery of the apoproteins in SP and PL. This fraction was therefore supplemented with protein purified separately to attain the initial levels present in CLSE. Biochemical analyses confirmed that the resulting preparations had the expected composition not only of protein, neutral lipids and phospholipids, but also of the phospholipid head groups. In addition to these fractions obtained with acidified solvent, elution of CLSE with chloroform-methanol without acid yielded the zwitterionic phospholipids substantially depleted of anionic phosphatidylglycerol and phosphatidylinositol. Limited interfacial measurements also demonstrated that the process of separation did not alter the fundamental surface characteristics of the surfactant constituents. Recombined CLSE (rCLSE) reconstituted from all of the separated components had surface activity indistinguishable from the original CLSE. The individual fractions of surfactant components also had average molecular areas at the air-liquid interface which agreed with predictions based on their biochemical composition. These well defined preparations of the hydrophobic constituents of pulmonary surfactant provide the basis for future studies to establish the role of individual components in the function of this complex surface active material.

Original languageEnglish (US)
Pages (from-to)1386-1394
Number of pages9
JournalJournal of Lipid Research
Volume35
Issue number8
StatePublished - 1994

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Surface-Active Agents
Lung
Phospholipids
Chloroform
Methanol
Proteins
Lipids
Pulmonary Surfactants
Phosphatidylglycerols
Apoproteins
Acids
Gel permeation chromatography
Phosphatidylinositols
Protein C
Chemical analysis
Gel Chromatography
Air
Recovery
Degradation
Liquids

Keywords

  • proteolipids
  • pulmonary surfactant
  • surface active agents
  • surfactant apoproteins
  • surfactant composition
  • surfactant phospholipids

ASJC Scopus subject areas

  • Endocrinology

Cite this

Separation of subfractions of the hydrophobic components of calf lung surfactant. / Hall, Stephen (Steve); Wang, Z.; Notter, R. H.

In: Journal of Lipid Research, Vol. 35, No. 8, 1994, p. 1386-1394.

Research output: Contribution to journalArticle

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