Interaction of surfactant protein A with cellular myosin.
ABSTRACT The goal of the current investigation was to characterize, purify, and identify the proteins that bind surfactant protein A (SP-A). Several polypeptides were purified by SP-A affinity chromatography, and the 200 kD major polypeptide that reacted with SP-A on ligand blots was purified further by preparative SDS-PAGE. Protein sequencing of proteolytically derived subfragments of this polypeptide gave sequences that corresponded completely with nonmuscle (cellular) myosin heavy chain. The 200 kD polypeptide was then found to be immunoreactive with antibodies against cellular myosin. A smaller polypeptide of 135 kD also binds SP-A and appears to be a proteolytic fragment of the 200 kD peptide. The ability of SP-A to bind myosin was confirmed in a microtiter well assay and was found to be concentration dependent. We speculated that the physiologic relevance of the interaction of SP-A with myosin might be to facilitate clearance of myosin from the alveolar subphase following its release during lung injury. In support of this hypothesis, we found that there were detectable levels of myosin in lavage fluid and that SP-A could indeed enhance uptake and degradation of myosin by alveolar macrophages.
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ABSTRACT: Surfactant protein A (SP-A) has been shown to bind to and regulate the functions of both alveolar type II cells and immune cells including alveolar macrophages. The interaction of SP-A with type II cells has been shown in vitro to inhibit lipid secretion and to promote the uptake of lipid by these cells and these observations led to the hypothesis that SP-A plays an important role in regulating surfactant turnover and metabolism. The finding that mice made deficient in SP-A by homologous recombination (SP-A -/- mice) have relatively normal surfactant pool sizes has raised the possibility that either redundant mechanisms function in vivo to keep pool sizes normal in the absence of SP-A or that the in vitro findings are not significant in the context of the whole, unstressed animal. The interaction of SP-A with immune cells has been shown to affect a variety of responses which, in general, function to promote host defense against infection. Although SP-A receptors have been identified, additional studies will be required to elucidate the mechanism of interaction of SP-A with these cells and the relative importance of the different receptors in SP-A mediated regulation of cell function.Biochimica et Biophysica Acta 12/1998; 1408(2-3):241-63. DOI:10.1016/S0925-4439(98)00071-4 · 4.66 Impact Factor
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ABSTRACT: Surfactant protein A (SP-A) is a highly ordered, oligomeric glycoprotein that is secreted into the airspaces of the lung by the pulmonary epithelium. The in vitro activities of protein suggest diverse roles in pulmonary host defense and surfactant homeostasis, structure and surface activity. Functional mapping of SP-A using directed mutagenesis has identified domains which interact with surfactant phospholipids, alveolar type II cells and microbes. Recently developed genetically manipulated animal models are beginning to clarify the critical physiological roles for SP-A in the normal lung, and in the pathophysiology of pulmonary disease.Biochimica et Biophysica Acta 12/1998; 1408(2-3):109-31. DOI:10.1016/S0925-4439(98)00062-3 · 4.66 Impact Factor
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ABSTRACT: Glycoprotein-340 (gp-340) was first identified as a surfactant protein (SP)-D-binding molecule purified from lung lavage of patients with alveolar proteinosis (Holmskov, et al., J. Biol. Chem. 1997;272:13743). In purifying SP-A from proteinosis lavage, we isolated a protein that copurifies with SP-A and SP-D and that was later found by protein sequencing to be gp-340. We have shown that soluble gp-340 binds SP-A in a calcium-dependent manner independent of the lectin activity of SP-A. To examine the functional significance of this interaction, we tested the ability of soluble gp-340 to block SP-A binding to and stimulation of the chemotaxis of alveolar macrophages. We found that gp-340 does not affect the binding of SP-A to alveolar macrophages over a wide range of SP-A concentrations, nor does it inhibit the ability of SP-A to stimulate macrophage chemotaxis. We also found that gp-340 alone stimulates the random migration (chemokinesis) of alveolar macrophages in a manner independent of SP-A-stimulated chemotaxis. These results suggest that gp-340 is not a cell-surface receptor necessary for SP-A stimulation of chemotaxis, and show that gp-340 can directly affect macrophage function.American Journal of Respiratory Cell and Molecular Biology 05/1999; 20(4):759-68. DOI:10.1165/ajrcmb.20.4.3439 · 3.99 Impact Factor