Abstract
Class-I restricted T cell-associated molecule (CRTAM) is a member of the immunoglobulin superfamily, and it is closely related to nectin-like protein. CRTAM is expressed in activated CD8 T cells, NKT cells, NK cells and in a subpopulation CD4 T cells. In this study, we produce as recombinant proteins, the Ig-domains of CRTAM (IgV-IgC), the IgV, and the IgC. These proteins were successfully purified in the soluble fraction only if the stalk region was included. The recombinant CRTAM recognizes its ligand nectin-like 2 in a cell-free system. We also demonstrate that the IgC domain of CRTAM is recognized by the anti-hCRTAM monoclonal antibody C8 with a 0.62 nM affinity. In conclusion, the stalk region of CRTAM provides solubility for the expression of its Ig-domains as recombinant proteins.
Keywords:
CRTAM; Nectin-like 2; Recombinant protein; Superfamily of immunoglobulins.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Antibodies, Monoclonal / biosynthesis
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Antibodies, Monoclonal / chemistry
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Binding Sites
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Cell Adhesion Molecule-1 / genetics*
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Cell Adhesion Molecule-1 / immunology
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Cell Adhesion Molecule-1 / metabolism
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Cell-Free System / chemistry*
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Cloning, Molecular
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Escherichia coli / genetics
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Escherichia coli / metabolism
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Female
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Gene Expression
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Genetic Vectors / chemistry
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Genetic Vectors / metabolism
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Humans
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Hybridomas / chemistry
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Immunoglobulin Domains / genetics*
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Immunoglobulins / genetics*
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Immunoglobulins / immunology
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Immunoglobulins / metabolism
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Jurkat Cells
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Mice
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Mice, Inbred BALB C
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Protein Binding
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Recombinant Proteins / genetics
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Recombinant Proteins / immunology
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Recombinant Proteins / metabolism
Substances
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Antibodies, Monoclonal
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CADM1 protein, human
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Cell Adhesion Molecule-1
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Immunoglobulins
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Recombinant Proteins
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class-I restricted T cell-associated molecule