Construction of Hybrid Protein Combining Immunoglobulin and Epidermal Growth Factor Receptor and Its Expression on the Myeloma Cell Membrane.

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  • 新しい免疫センサーのためのＩｇＭ‐上皮増殖因子受容体融合タンパク質の試作とそのミエローマ細胞における膜発現
  • アタラシイ メンエキ センサー ノ タメ ノ IgM ジョウヒ ゾウショク イ

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Abstract

A hybrid receptor gene that combines the murine immunoglobulin IgM gene and a cDNA encoding the cytoplasmic domain of human epidermal growth factor receptor (EGFR) was constructed to cultivate the possibility of a new biosensor. The construct was transfected into myeloma cell which expresses Ig light chain. After selection, transfectant cells which express IgM-EGFR chimeric receptor protein were obtained.<BR>The chimeric protein could assemble to H2L2 heterotetramer conformation and had the same antigen 4-Hydroxy-3-Nitrophenacetyl binding activity as its parent murine IgM. While wild-type membrane IgM is not transported to the plasmamembrane of this cell line, the chimeric receptor was transported from endoplasmic reticulum to the plasma membrane through Golgi apparatus. Moreover, this antigen receptor had in vitro protein tyrosine kinase activity, which was necessary for intracellular signal transduction. The results indicate that myeloma cells can be used as an expression system of such biosensor proteins.

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