Identification of a putative surfactant convertase in rat lung as a secreted serine carboxylesterase

In the alveolar lumen, pulmonary surfactant converts from the contents of secreted lamellar bodies to tubular myelin to apoprotein-depleted vesicles during respiration. Using an in vitro system, researchers have reported that the conversion of tubular myelin to vesicles is blocked by inhibitors of serine hydrolase activity and have tentatively ascribed "convertase" activity to a diisopropyl fluorophosphate (DFP)-binding protein in mouse bronchoalveolar lavage (BAL). We purified and sequenced the homologous enzyme from rat BAL fluid. Amino acid sequence from the amino terminus and an internal cyanogen bromide peptide of the purified rat DFP-binding protein perfectly match the sequence of the carboxylesterase ES-2. Although ES-2 was initially cloned from liver, we found a 1.8-kilobase mRNA for ES-2 in decreasing relative abundance in rat liver, kidney, and lung but not in heart or spleen. Although further studies are required to establish the identity between "convertase" and ES-2 or a homologous member of the carboxylesterase family, our results raise the possibility that a protein with esterase/lipase activity plays a role in extracellular surfactant metabolism

Kinetics of phospholipid membrane fusion induced by surfactant apoproteins A and B

AbstractSurfactant apoproteins A (SP-A) and B (SP-B) interact with the lipids of surfactant and such protein-lipid interactions may be of importance in several of the steps in the surfactant cycle. We analyzed the kinetics of fusion of dipalmitoylphosphatidylcholine-phosphatidylglycerol (DPPC:PG; 7:3, w/w) phospholipid vesicles induced by SP-B alone, in the presence of 5 mM calcium, and in the presence of calcium and SP-A. Membrane fusion was measured by the method of resonance energy transfer between non-exchangeable fluorophores incorporated in the membrane. Data were analyzed using a mass action kinetic model for membrane fusion between phospholipid vesicles. We found a SP-B dose-dependent increase in lipid mixing within a range of phospholipid concentration of 5 to 100 μM. Calcium caused a small additive increase in lipid mixing, but calcium and SP-A combined markedly increased lipid mixing induced by SP-B. Both aggregation and fusion rate constants increased with an increase in the SP-B/lipid ratio. In the presence of calcium and SP-A, the number of vesicles per fusion product markedly increased, as did the aggregation rate constants, whereas the fusion rate constants remained essentially unchanged

Protective Role of Surfactant Protein D in Ocular Staphylococcus aureus Infection.

Staphylococcus aureus is one of the most common pathogens causing keratitis. Surfactant protein D (SP-D) plays a critical role in host defense and innate immunity. In order to investigate the role of SP-D in ocular S. aureus infection, the eyes of wild-type (WT) and SP-D knockout (SP-D KO) C57BL/6 mice were infected with S. aureus (10(7) CFU/eye) in the presence and absence of cysteine protease inhibitor(E64).Bacterial counts in the ocular surface were examined 3, 6, 12, 24 hrs after infection. Bacterial phagocytosis by neutrophils and bacterial invasion in ocular epithelial cells were evaluated quantitatively. S. aureus-induced ocular injury was determined with corneal fluorescein staining. The results demonstrated that SP-D is expressed in ocular surface epithelium and the lacrimal gland; WT mice had increased clearance of S. aureus from the ocular surface (p<0.05) and reduced ocular injury compared with SP-D KO mice. The protective effects of SP-D include increased bacterial phagocytosis by neutrophils (p<0.05) and decreased bacterial invasion into epithelial cells (p<0.05) in WT mice compared to in SP-D KO mice. In the presence of inhibitor (E64), WT mice showed enhanced bacterial clearance (p<0.05) and reduced ocular injury compared to absent E64 while SP-D KO mice did not. Collectively, we concluded that SP-D protects the ocular surface from S. aureus infection but cysteine protease impairs SP-D function in this murine model, and that cysteine protease inhibitor may be a potential therapeutic agent in S. aureus keratitis

Protective role of surfactant protein D in ocular staphylococcus aureus infection

© 2015 Zhang et al.Staphylococcus aureus is one of the most common pathogens causing keratitis. Surfactant protein D (SP-D) plays a critical role in host defense and innate immunity. In order to investigate the role of SP-D in ocular S. aureus infection,

Interactions of The Low Molecular Weight Group of Surfactant-associated Proteins (sp 5–18) With Pulmonary Surfactant Lipids

The interaction of the low molecular weight group of surfactant-associated proteins, SP 5–18, with the major phospholipids of pulmonary surfactant was studied by fluorescence measurements of liposomal permeability and fusion, morphological studies, and surface activity measurements. The ability of SP 5–18 to increase the permeability of large unilamellar lipid vesicles was enhanced by the presence of negatively charged phospholipid. The permeability of these vesicles increased as the protein concentration was raised and the pH was lowered. SP 5–18 also induced leakage from liposomes made both from a synthetic surfactant lipid mixture and from lipids separated from SP 5–18 during its purification from canine sources. When SP 5–18 was added to egg phosphatidylglycerol liposomes, the population of liposomes which became permeable leaked all encapsulated contents, while the remaining liposomes did not leak at all. The extent of leakage was higher in the presence of 3 mM calcium. SP 5–18 also induced lipid mixing between two populations of egg phosphatidylglycerol liposomes in the presence of 3 mM calcium, as monitored by resonance energy transfer between two different fluorescent lipid probes, N-(7-nitro-2,l,3-benzoxadiazol-4-yl)phosphatidylethanolamine and N-(lissamine rhodamine B sulfonyl)phosphatidylethanolamine. Negative-staining electron microscopy showed that the addition of SP 5–18 and 3 mM calcium produced vesicles twice the size of control egg phosphatidylglycerol liposomes. In addition, surface balance measurements revealed that the adsorption of liposomal lipids to an air/water interface was enhanced by the presence of SP 5–18, negatively charged phospholipids, and 3 mM calcium. These observations suggest a similar lipid dependence for the interactions observed in the fluorescence and adsorption experiments. © 1988, American Chemical Society. All rights reserved

Surfactant protein D binds genomic DNA and apoptotic cells, and enhances their clearance, in vivo

Collectins, such as surfactant protein D (SP-D), bind apoptotic cells; however, the ligands that they recognize on these cells are unknown. We hypothesized that SP-D binds to the DNA present on these cells. We show that SP-D binds and aggregates mouse alveolar macrophage DNA effectively. Alveolar macrophages of SP-D((-/-)) mice contained more nicked DNA than those of SP-A((-/-)) and wild type mice. Our results also suggest that carbohydrate recognition domains of SP-D may recognize DNA present on the apoptotic cells. Therefore, cell-surface DNA could be a ligand for recognition of apoptotic cells by collectins