scholarly journals Surface properties, morphology and protein composition of pulmonary surfactant subtypes

1996 ◽  
Vol 320 (2) ◽  
pp. 599-605 ◽  
Author(s):  
Esther PUTMAN ◽  
Lambert A. J. M. CREUWELS ◽  
Lambert M. G. van GOLDE ◽  
Henk P. HAAGSMAN
Keyword(s):  
Pulmonary Surfactant ◽  
Structural Organization ◽  
Adsorption Process ◽  
Protein A ◽  
Protein Composition ◽  
High Adsorption ◽  
Adsorption Studies ◽  
Natural Surfactants ◽  
Surfactant Metabolism

Separation of surfactant subtypes is now commonly used as a parameter in assessing the amount of active compared with inactive material in various models of lung injury. The protein content, morphology and surface activity were determined of the heavy and light subtype isolated by differential centrifugation. Here we report the presence of surfactant proteins B and C in the heavy subtype but not in the light subtype. Adsorption studies revealed that separation of fast adsorbing bronchoalveolar lavage resulted in slowly adsorbing heavy and light subtypes. Surfactant, reconstituted from heavy and light fractions, did not show a high adsorption rate. It is concluded that the isolation procedures might result in a loss of fast adsorbing surfactant structures. Surface area cycling was used as a model in vitro for the extracellular surfactant metabolism. The heavy subtype is converted into the light subtype during conversion. Conversion performed with resuspended heavy subtype revealed the generation of a disparate subtype. Furthermore it was found that the conversion was dependent on preparation and handling of the samples before cycling. Finally, adsorption studies at low surfactant concentrations revealed a delayed adsorption of lipid-extracted surfactants compared with natural surfactants. These observations emphasize the importance of the (surfactant-associated protein A-dependent) structural organization of surfactant lipids in the adsorption process.

Surfactant metabolism in surfactant protein A-deficient mice

1997 ◽  
Vol 272 (3) ◽  
pp. L479-L485 ◽  
Author(s):  
M. Ikegami ◽  
T. R. Korfhagen ◽  
M. D. Bruno ◽  
J. A. Whitsett ◽  
A. H. Jobe
Keyword(s):  
Lung Tissue ◽  
Protein A ◽  
Surfactant Protein ◽  
Normal Lung ◽  
Tissue Slices ◽  
Normal Lung Function ◽  
Deficient Mice ◽  
Surfactant Metabolism

In the present study we asked if surfactant metabolism was altered in surfactant protein (SP) A-deficient mice in vivo. Although previous studies in vitro demonstrated that SP-A modulates surfactant secretion and reuptake by type II cells, mice made SP-A deficient by homologous recombination grow and reproduce normally and have normal lung function. Alveolar and lung tissue saturated phophatidylcholine (Sat PC) pools were 50 and 26% larger, respectively, in SP-A(-/-) mice than in SP-A(+/+) mice. Radiolabeled choline and palmitate incorporation into lung Sat PC was similar both in vivo and for lung tissue slices in vitro from SP-A(+/+) and SP-A(-/-) mice. Percent secretion of radiolabeled Sat PC was unchanged from 3 to 15 h, although SP-A(-/-) mice retained more labeled Sat PC in the alveolar lavages at 48 h (consistent with the increased surfactant pool sizes). Clearance of radiolabeled dipalmitoylphosphatidylcholine and SP-B from the air spaces after intratracheal injection was similar in SP-A(-/-) and SP-A(+/+) mice. Lack of SP-A had minimal effects on the overall metabolism of Sat PC or SP-B in mice.

scholarly journals Surfactant protein A inhibits T cell proliferation via its collagen-like tail and a 210-kDa receptor

1998 ◽  
Vol 275 (4) ◽  
pp. L679-L686 ◽  
Author(s):  
Paul Borron ◽  
Francis X. McCormack ◽  
Baher M. Elhalwagi ◽  
Zissis C. Chroneos ◽  
James F. Lewis ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
T Cell ◽  
T Lymphocyte ◽  
Lymphocyte Proliferation ◽  
Pulmonary Surfactant ◽  
Protein A ◽  
T Cell Proliferation ◽  
T Lymphocyte Proliferation ◽  
Human T Lymphocyte

Investigation of possible mechanisms to describe the hyporesponsiveness of pulmonary leukocytes has led to the study of pulmonary surfactant and its constituents as immune suppressive agents. Pulmonary surfactant is a phospholipid-protein mixture that reduces surface tension in the lung and prevents collapse of the alveoli. The most abundant protein in this mixture is a hydrophilic molecule termed surfactant-associated protein A (SP-A). Previously, we showed that bovine (b) SP-A can inhibit human T lymphocyte proliferation and interleukin-2 production in vitro. Results presented in this investigation showed that different sources of human SP-A and bSP-A as well as recombinant rat SP-A inhibited human T lymphocyte proliferation in a dose-dependent manner. A structurally similar collagenous protein, C1q, did not block the in vitro inhibitory action of SP-A. The addition of large concentrations of mannan to SP-A-treated cultures also did not disrupt inhibition, suggesting that the effect is not mediated by the carbohydrate recognition domain of SP-A. Use of recombinant mutant SP-As revealed that a 36-amino acid Arg-Gly-Asp (RGD) motif-containing span of the collagen-like domain was responsible for the inhibition of T cell proliferation. A polyclonal antiserum directed against an SP-A receptor (SP-R210) completely blocked the inhibition of T cell proliferation by SP-A. These results emphasize a potential role for SP-A in dampening lymphocyte responses to exogenous stimuli. The data also provide further support for the concept that SP-A maintains a balance between the clearance of inhaled pathogens and protection against collateral immune-mediated damage.

Pulmonary surfactant protein-A (SP-A) modulates human PBMC proliferation and cytokine production in vitro

Cytokine ◽  
1994 ◽  
Vol 6 (5) ◽  
pp. 539
Author(s):  
Paul Borron ◽  
L.J. Fraher ◽  
R.G. McFadden ◽  
J.F. Lewis ◽  
F. Possmayer
Keyword(s):  
Pulmonary Surfactant ◽  
Cytokine Production ◽  
Protein A ◽  
Surfactant Protein ◽  
Surfactant Protein A ◽  
Human Pbmc ◽  
Pulmonary Surfactant Protein

Surfactant protein A regulates surfactant phospholipid clearance after LPS-induced injury in vivo

2002 ◽  
Vol 283 (1) ◽  
pp. L76-L85 ◽  
Author(s):  
Omar A. Quintero ◽  
Thomas R. Korfhagen ◽  
Jo Rae Wright
Keyword(s):  
Acute Lung Injury ◽  
Lung Injury ◽  
Pulmonary Surfactant ◽  
Protein A ◽  
Surfactant Protein ◽  
Surfactant Protein A ◽  
Wild Type ◽  
Surfactant Phospholipids

Previous in vitro studies have suggested that surfactant protein A (SP-A) may play a role in pulmonary surfactant homeostasis by mediating surfactant secretion and clearance. However, mice made deficient in SP-A [SP-A (−/−) animals] have relatively normal levels of surfactant compared with wild-type SP-A (+/+) animals. We hypothesize that SP-A may play a role in surfactant homeostasis after acute lung injury. Bacterial lipopolysaccharide was instilled into the lungs of SP-A (−/−) mice and SP-A (+/+) mice to induce injury. Surfactant phospholipid levels were increased 1.6-fold in injured SP-A (−/−) animals, although injury did not alter [3H]choline or [14C]palmitate incorporation into dipalmitoylphosphatidylcholine (DPPC), suggesting no change in surfactant synthesis/secretion 12 h after injury. Clearance of [3H]DPPC from the lungs of injured SP-A (−/−) animals was decreased by ∼40%. Instillation of 50 μg of exogenous SP-A rescued both the clearance defect and the increased phospholipid defect in injured SP-A (−/−) animals, suggesting that SP-A may play a role in regulating clearance of surfactant phospholipids after acute lung injury.

Inhaled nitric oxide injures the pulmonary surfactant system of lambs in vivo

1996 ◽  
Vol 270 (2) ◽  
pp. L273-L280 ◽  
Author(s):  
S. Matalon ◽  
V. DeMarco ◽  
I. Y. Haddad ◽  
C. Myles ◽  
J. W. Skimming ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Bronchoalveolar Lavage ◽  
Surface Properties ◽  
Pulmonary Surfactant ◽  
Protein A ◽  
Surfactant Protein ◽  
Surfactant System ◽  
Minimum Surface Tension

Nitric oxide (.NO) is a free radical, and as such may damage the pulmonary surfactant system. To determine the potential toxicity of .NO in vivo, we exposed 35 newborn lambs to 0, 20, 80 or 200 ppm .NO in either 21 or 60% O2 for 6 h. At the end of the exposure, lambs had normal values of arterial Po2, Pco2, and pH; total protein concentration in the bronchoalveolar lavage was also at normal levels. There were no differences in the surface properties of surfactant among the air or 60% O2 groups. Pulmonary surfactant samples, isolated from the bronchoalveolar lavage of lambs breathing air or 20 ppm .NO and reconstituted at a lipid concentration of 3 mg/ml, reached a low minimum surface tension (Tmin < 3 mN/m) in a pulsating bubble surfactometer. On the other hand, abnormal surface properties were observed in 36 and 60% of surfactant samples isolated from lungs of lambs that breathed 80 or 200 ppm .NO, respectively. These findings were confirmed using a captive bubble surfactometer. Surfactant protein A, isolated from the lungs of lambs that breathed 200 ppm .NO, exhibited decreased ability to aggregate lipids in vitro. These data are consistent with injury to the surfactant apoproteins during inhalation of either 80 or 200 ppm .NO for 6 h.

Preferential uptake of small-aggregate fraction of pulmonary surfactant in vitro

1997 ◽  
Vol 273 (2) ◽  
pp. L468-L477 ◽  
Author(s):  
A. D. Horowitz ◽  
K. Kurak ◽  
B. Moussavian ◽  
J. A. Whitsett ◽  
S. E. Wert ◽  
...  
Keyword(s):  
Pulmonary Surfactant ◽  
Protein A ◽  
Surfactant Protein ◽  
Surfactant Protein A ◽  
Epifluorescence Microscopy ◽  
Mouse Lung ◽  
Metabolic Clearance ◽  
Small Aggregate ◽  
The Difference

Homeostasis of pulmonary surfactant requires metabolic clearance of surfactant forms with decreased surface activity. Rabbit pulmonary surfactant was labeled in vivo with rhodamine-labeled dipalmitoylphosphatidylethanolamine (R-DPPE), isolated, and fractionated into large- and small-aggregate subfractions by differential centrifugation. Endocytosis of large (LA)- and small (SA)-aggregate surfactant by a mouse lung epithelial cell line (MLE-12) was evaluated in vitro by epifluorescence microscopy. More SA than LA surfactant was taken up by MLE-12 cells. Endocytosis of SA and LA surfactant was inhibited by preincubation of the subfractions with surfactant protein A and 3.3 mM Ca2+. The difference in uptake between SA and LA surfactant was lost for reconstituted organic extracts of the subfractions. Much of the difference in uptake of SA and LA surfactant may be attributed to the greater concentration of surfactant protein A in LA surfactant.

Mercury Removal from Wastewater by Steamed Hoof Powder

1999 ◽  
Vol 40 (7) ◽  
pp. 109-116 ◽  
Author(s):  
M. H. Ansari ◽  
A. M. Deshkar ◽  
P. S. Kelkar ◽  
D. M. Dharmadhikari ◽  
M. Z. Hasan ◽  
...  
Keyword(s):  
Heavy Metals ◽  
Adsorption Capacity ◽  
Adsorption Process ◽  
Adsorption Equilibrium ◽  
Light Metal ◽  
Mercury Removal ◽  
High Adsorption ◽  
Surface Sites ◽  
Ph Values ◽  
High Adsorption Capacity

Steamed Hoof Powder (SHP), size &lt; 53μ, was observed to have high adsorption capacity for Hg(II) with &gt;95% removal from a solution containing 100 mg/L of Hg(II) with only 0.1% (W/V) concentration of SHP. The SHP has good settling properties and gives clear and odour free effluent. Studies indicate that pH values between 2 and 10 have no effect on the adsorption of Hg(II) on SHP. Light metal ions like Na+, K+, Ca2+ and Mg2+ up to concentrations of 500 mg/L and heavy metals like Cu2+, Zn2+, Cd2+, Co2+, Pb2+, Ni2+, Mn2+, Cr3+, Cr6+, Fe2+ and Fe3+ up to concentrations of 100 mg/L do not interfere with the adsorption process. Anions like sulphate, acetate and phosphate up to concentrations of 200 mg/L do not interfere. Chloride interferes in the adsorption process when Hg(II) concentration is above 9.7 mg/L. The adsorption equilibrium was established within two hours. Studies indicate that adsorption occurs on the surface sites of the adsorbent.

Proteomics-Based Characterization of the Effects of MMP14 on the Protein Content of Exosomes from Corneal Fibroblasts

2020 ◽  
Vol 27 (10) ◽  
pp. 979-988
Author(s):  
Kyu-Yeon Han ◽  
Jin-Hong Chang ◽  
Dimitri T. Azar
Keyword(s):  
Protein Content ◽  
Catalytic Domain ◽  
Protein Composition ◽  
Proteomics Analysis ◽  
Knock Out ◽  
Corneal Fibroblast ◽  
Flow Cytometric ◽  
Corneal Fibroblasts ◽  
Liquid Chromatography Tandem Mass

Background: Exosomes secreted by corneal fibroblasts contain matrix metalloproteinase (MMP) 14, which is known to influence pro-MMP2 accumulation on exosomes. Accordingly, we hypothesized that the enzymatic activity of MMP14 may alter the protein content of corneal fibroblast- secreted exosomes. Objective: The aim of this study was to investigate the effects of MMP14 on the composition and biological activity of corneal fibroblast-derived exosomes. Methods: Knock out of the catalytic domain (ΔExon4) of MMP14 in corneal fibroblasts was used to determine the effect of MMP14 expression on the characteristics of fibroblast-secreted exosomes. The amount of secreted proteins and their size distribution were measured using Nano Tracking Analysis. Proteins within exosomes from wild-type (WT) and ΔExon4-deficient fibroblasts were identified by liquid chromatography-tandem mass spectrometry (MS/MS) proteomics analysis. The proteolytic effects of MMP14 were evaluated in vitro via MS identification of eliminated proteins. The biological functions of MMP14-carrying exosomes were investigated via fusion to endothelial cells and flow cytometric assays. Results: Exosomes isolated from WT and ΔExon4-deficient fibroblasts exhibited similar size distributions and morphologies, although WT fibroblasts secreted a greater amount of exosomes. The protein content, however, was higher in ΔExon4-deficient fibroblast-derived exosomes than in WT fibroblast-derived exosomes. Proteomics analysis revealed that WT-derived exosomes included proteins that regulated cell migration, and ΔExon4 fibroblast-derived exosomes contained additional proteins that were cleaved by MMP14. Conclusion: Our findings suggest that MMP14 expression influences the protein composition of exosomes secreted by corneal fibroblasts, and through those biological components, MMP14 in corneal fibroblasts derived-exosomes may regulate corneal angiogenesis.

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