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(Received for publication, October 5, 1995, and in revised form, April 11, 1996)
From the In the present report we have characterized the
binding of surfactant protein A (SP-A) to bone marrow-derived
macrophages, U937 cells, alveolar macrophages, and type II epithelial
cells. The binding of SP-A to all cell types is
Ca2+-dependent and trypsin-sensitive, but type
II cells express distinct Ca2+-independent binding sites.
The binding of SP-A to macrophages is independent of known cell surface
carbohydrate-specific receptors and of glycoconjugate binding sites on
the surface of the cells and is distinct from binding to C1q receptors.
Based on ligand blot analysis, both type II cells and macrophages
express a 210-kDa SP-A-binding protein. The 210-kDa protein was
purified to apparent homogeneity from U937 macrophage membranes using
affinity chromatography with noncovalently immobilized surfactant
protein A, and was purified from rat lung by differential detergent and
salt extraction of isolated rat lung membranes. Polyclonal antibodies
against the rat lung SP-A-binding protein inhibit binding of SP-A to
both type II cells and macrophages, indicating that the 210-kDa protein
is expressed on the cell surface. The polyclonal antibodies also block
the SP-A-mediated inhibition of phospholipid secretion by type II
cells, indicating that the 210-kDa protein is a functional cell-surface
receptor on type II cells. In a separate report we have determined that
antibodies to the SP-A receptor block the SP-A-mediated uptake of
Mycobacterium bovis, indicating that the macrophage SP-A
receptor is involved in SP-A-mediated clearance of pathogens.
Volume 271, Number 27,
Issue of July 5, 1996
pp. 16375-16383
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.
,
,
and
''
Division of Pulmonary Biology, Children's
Hospital Medical Center, Cincinnati, Ohio 45229-3039; Departments
of
Biochemistry, ¶ Medicine, and '' Pathology, Vanderbilt
University, Nashville, Tennessee 37232; and the
Department of Veteran's Affairs Medical
Center, Nashville, Tennessee 37203
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