A truncated recombinant intercellular adhesion molecule-1 inhibits adhesion of leukemic cell lines to upregulated endothelial cells.
Intercellular adhesion molecule-1, a member of the immunoglobulin supergene family, is the ligand for the integrin lymphocyte function associated antigen-1. Intercellular adhesion molecule-1 and lymphocyte function associated antigen-1 binding interactions mediate leukocyte adherence and migration. Previous work has shown that the adherence of lymphocyte function associated antigen-1 is directed to the first immunoglobulinlike domain of the endothelial cell surface protein intracellular adhesion molecule-1. We have constructed a truncated intercellular adhesion molecule-1 gene encoding the first 185 amino acids from the amino terminus and overexpressed it in Escherichia coli. The recombinant protein was purified from insoluble inclusion bodies and refolded into an active conformation by a denaturation/renaturation cycle. The identity of the protein was confirmed by microsequencing and by Western blot analysis using a polyclonal antibody to ICAM-1. We have demonstrated that this soluble region of the otherwise membrane-bound ligand is an inhibitor of Molt or HL-60 cell adhesion to cytokine-stimulated endothelial cells.
Ross
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Department of Cell Biology, Glaxo Inc. Research Institute, Research Triangle Park, North Carolina 27709.
Davis
C M
CM
Molony
L
L
D000595:Amino Acid Sequence; D001483:Base Sequence; D015153:Blotting, Western; D002448:Cell Adhesion; D004730:Endothelium, Vascular / Q000166:cytology; D004926:Escherichia coli; D005434:Flow Cytometry; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000737:chemistry* / Q000235:genetics / Q000494:pharmacology; D007938:Leukemia / Q000473:pathology*; D015473:Leukemia, Promyelocytic, Acute / Q000473:pathology; D016169:Lymphocyte Function-Associated Antigen-1 / Q000378:metabolism; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000737:chemistry / Q000494:pharmacology; D013329:Structure-Activity Relationship; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D014471:Umbilical Veins
D016169:Lymphocyte Function-Associated Antigen-1; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha; D018799:Intercellular Adhesion Molecule-1