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Am J Physiol Lung Cell Mol Physiol 277: L1179-L1189, 1999;
1040-0605/99 $5.00
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Vol. 277, Issue 6, L1179-L1189, December 1999

SP-B refining of pulmonary surfactant phospholipid films

Kaushik Nag1,2,3, James G. Munro4, Kevin Inchley1,2,3, Samuel Schürch5, Nils O. Petersen4, and Fred Possmayer1,2,3

1 Department of Obstetrics and Gynaecology, 2 Department of Biochemistry, 3 Medical Research Council Group in Fetal and Neonatal Health and Development, and 4 Department of Chemistry, University of Western Ontario, London, Ontario N6A 5A5; and 5 Respiratory Research Group, Faculty of Medicine, University of Calgary, Alberta, Canada T2N 4N1

Pulmonary surfactant stabilizes the alveoli by lining the air-fluid interface with films that reduce surface tension to near 0 mN/m (gamma min). Surfactant protein B (SP-B) enhances the surface activity of surfactant phospholipids. A captive bubble tensiometer (CBT) was used to study the properties of adsorbed films of dipalmitoylphosphatidylcholine (DPPC) with acidic 1-palmitoyl-2-oleoyl-sn-glycero-3-[phospho-rac-(1-glycerol)] (POPG) or neutral 1-palmitoyl-2-oleoyl-sn-glycerol-3-phosphocholine with (7:3) and without 1% dimeric SP-B. SP-B enhanced the adsorption rate of DPPC-containing neutral or acidic lipid suspensions (1 mg/ml) to a similar extent. Quasi-static cycling of these films revealed that SP-B significantly decreased the film area reduction required to reach gamma min for the acidic but not for the neutral system. The results obtained with DPPC-phosphatidylglycerol (PG)-SP-B were consistent with selective DPPC adsorption into the surface monolayer during film formation. Film area reduction required to reach gamma min with this system (with and without calcium) approached that of pure DPPC, suggesting selective DPPC insertion and PG squeeze-out. Dynamic cycling of such films showed that larger film area reductions were required to reach gamma min for the neutral than for acidic system, even after 20 cycles. Fluorescence microscopy of solvent-spread DPPC-POPG-SP-B planar films revealed highly condensed structures at ~25 mN/m, although no specific PG phase-segregated structures could be identified. The study suggests that specific interactions of SP-B with acidic phospholipids of surfactant may be involved in the generation and maintenance of DPPC-rich films in the alveoli.

captive bubble tensiometer; surface tension; dipalmitoylphosphatidylcholine; phosphatidylglycerol; lipid-protein interactions; surfactant protein B; respiratory distress syndrome; squeeze-out; selective adsorption


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