Functional studies of truncated soluble intercellular adhesion molecule 1 expressed in Escherichia coli.
We have expressed in Escherichia coli the two N-terminal immunoglobulin (Ig)-like domains of the intercellular adhesion molecule 1 (ICAM-1). The first 188 residues of ICAM-1 were expressed with an N-terminal methionine (MP188) or as a maltose-binding fusion protein which was cleaved with factor Xa (XP188). After refolding, both MP188 and XP188 were active in binding to the leukocyte integrin lymphocyte function-associated antigen 1, which has previously been shown to bind to the N-terminal Ig domain of ICAM-1. The major group of rhinoviruses and malaria-infected erythrocytes bind to distinct sites within the first Ig-like domain of ICAM-1. Both MP188 and XP188 bound to malaria-infected erythrocytes; however, only XP188 inhibited human rhinovirus plaque formation. A product (MdQ1P188) with the initiation methionine fused to residue 2, i.e., with glutamine 1 deleted, inhibited plaque formation. MdQ1P188 was able to induce a conformational change of the virus capsid as shown by conversion of 149S particles to 85S particles, whereas MP188 had no effect. These results show that functionally active fragments of ICAM-1 can be produced in E. coli, that glycosylation is not required for ligand binding, and that the N-terminal residue of ICAM-1 is proximal to or part of the human rhinovirus-binding site.
Martin
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Center for Blood Research, Boston, Massachusetts 02115.
Martin
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Staunton
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Springer
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D000818:Animals; D000911:Antibodies, Monoclonal; D000998:Antiviral Agents / Q000378:metabolism / Q000494:pharmacology; D001483:Base Sequence; D001665:Binding Sites; D016466:CHO Cells / Q000502:physiology; D015815:Cell Adhesion Molecules / Q000302:isolation & purification / Q000502:physiology*; D006224:Cricetinae; D004912:Erythrocytes / Q000378:metabolism / Q000382:microbiology; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D018799:Intercellular Adhesion Molecule-1; D016169:Lymphocyte Function-Associated Antigen-1 / Q000378:metabolism; D008288:Malaria / Q000097:blood; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000302:isolation & purification / Q000494:pharmacology / Q000502:physiology*; D017510:Protein Folding; D012229:Rhinovirus / Q000187:drug effects / Q000378:metabolism
D000911:Antibodies, Monoclonal; D000998:Antiviral Agents; D015815:Cell Adhesion Molecules; D016169:Lymphocyte Function-Associated Antigen-1; D010446:Peptide Fragments; D018799:Intercellular Adhesion Molecule-1