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Protein J ; 39(3): 224-231, 2020 06.
Article in English | MEDLINE | ID: mdl-32300914

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.


Subject(s)
Cell Adhesion Molecule-1/genetics , Cell-Free System/chemistry , Immunoglobulin Domains/genetics , Immunoglobulins/genetics , Animals , Antibodies, Monoclonal/biosynthesis , Antibodies, Monoclonal/chemistry , Binding Sites , Cell Adhesion Molecule-1/immunology , Cell Adhesion Molecule-1/metabolism , Cloning, Molecular , Escherichia coli/genetics , Escherichia coli/metabolism , Female , Gene Expression , Genetic Vectors/chemistry , Genetic Vectors/metabolism , Humans , Hybridomas/chemistry , Immunoglobulins/immunology , Immunoglobulins/metabolism , Jurkat Cells , Mice , Mice, Inbred BALB C , Protein Binding , Recombinant Proteins/genetics , Recombinant Proteins/immunology , Recombinant Proteins/metabolism
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