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J Biol Chem, Vol. 273, Issue 8, 4725-4733, February 20, 1998

Molecular Comparison of Soluble Intercellular Adhesion Molecule (sICAM)-1 and sICAM-3 Binding to Lymphocyte Function-associated Antigen-1

Joseph R. Woska Jr.Dagger , Maurice M. Morelock, Deborah Durham JeanfavreDagger , Gary O. CavinessDagger , Barbara-Jean BormannDagger , and Robert RothleinDagger

From the Dagger  Cell Adhesion Group, Department of Immunological Diseases, Boehringer Ingelheim Pharmaceuticals, Inc. and the  Biophysics Group, Department of Inflammatory Diseases, Boehringer Ingelheim Pharmaceuticals, Inc., Ridgefield, Connecticut 06877

The interactions of intercellular adhesion molecules-1 and -3 (ICAM-1 and ICAM-3) with lymphocyte function-associated antigen-1 (LFA-1) have been characterized and compared on the molecular and cellular level. Enzyme-linked immunosorbent-based molecular assays have been utilized to calculate the binding affinities of soluble ICAM-1 (sICAM-1) and soluble ICAM-3 (sICAM-3) for LFA-1. Consistent with previously published data, we found that sICAM-1 binds to LFA-1 with an affinity of ~60 nM. In contrast, sICAM-3 binds to LFA-1 with an affinity ~9 times weaker (~550 nM). Both sICAM-1 and sICAM-3 require divalent cations for binding. Specifically, both Mg2+ and Mn2+ support high affinity adhesion, although interestingly, high concentrations of Ca2+ decrease the affinity of each molecule for LFA-1 substantially. Furthermore, a panel of anti-LFA-1 monoclonal antibodies were characterized for their ability to block sICAM-1 and sICAM-3/LFA-1 interactions in molecular and cellular assays to help distinguish binding sites on LFA-1 for both molecules. Finally, molecular and cellular competition experiments demonstrate that sICAM-1 and sICAM-3 compete with each other for binding to LFA-1. The above data demonstrate that sICAM-1 and sICAM-3 share a common binding site or an overlapping binding site on LFA-1 and that the apparent differences in binding sites can be attributed to different affinities of sICAM-1 and sICAM-3 for LFA-1.

Copyright © 1998 by The American Society for Biochemistry and Molecular Biology, Inc.
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