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(Received for publication, November 29,
1994; and in revised form, December 22, 1994) CD14 is a 55-kDa glycoprotein which binds lipopolysaccharide
(LPS) and enables LPS-dependent responses in a variety of cells. Recent
limited proteolysis studies have implicated a region in CD14 between
amino acids 57 and 64 as being involved in LPS interaction. To
specifically assess the importance of this region with respect to LPS
binding, we constructed a mutant sCD14
(sCD14
Volume 270,
Number 10,
Issue of March 10, 1995 pp. 5219-5224
©1995 by The American Society for Biochemistry and Molecular Biology, Inc.
) lacking amino acids 57-64.
sCD14 was isolated from the serum-free
conditioned medium of this cell line, and, in all assays, the purified
protein failed to recognize LPS or enable LPS-dependent responses in
cells. We also demonstrated that the region between amino acids 57 and
64 is required for binding of a neutralizing CD14 mAb, MEM-18. Native
polyacrylamide gel electrophoresis assays were used to demonstrate that
MEM-18 and LPS compete for the same binding site on CD14. These data
strongly suggest that the region spanning amino acids 57-64 binds
LPS and that formation of sCD14
LPS complex is required in order
for sCD14-mediated responses to occur.
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