G Protein .GAMMA. Subunits Coimmunoprecipitated with Antibodies against .ALPHA. Subunits: Identification of Major Isoforms in Cultured Cells by Silver Stain and Immunoblotting with Conventional Transfer Procedure.

概要

論文の詳細を見る
The βγ subunits of G proteins were coimmunoprecipitated with antibodies against various α subunits, and analyzed by silver stain and immunoblotting with conventional transfer procedure and membrane-blocking buffer containing 2% BSA. Multiple isoforms of γ were coimmunoprecipitated with no significant difference in form or ratio among the antibodies against α subunits used, suggesting antibodies against any α subunit could coimmuno-precipitate all forms of γ. Therefore, this method was applicable to analyze γ subunits in various cells, especially to clarify what forms of γ subunits are major components. The major isoforms were: γ5 in C6, NG108-15, HeLa, HEK293, and F9 cells; γ12 in Swiss 3T3 and BRL-3A cells; and γ3 in PC12 cells. In addition to most γ subunits identified, unidentified γ subunits were present in PC12, NG108-15, and BRL-3A cells. Furthermore, the method was applied to examine changes of isoforms of γ during differentiation of HL-60 cells. Undifferentiated cells mainly contained γ5, but retinoic acid treatment of cells replaced most γ5 with γ2. Thus, this method is useful to determine the major isoforms which seem to be the more important in cells.
社団法人日本生化学会の論文