Assays of recombinant adenylyl cyclases expressed in Sf9 cells.
This chapter outlines procedures for the expression of mammalian membrane-bound adenylyl cyclases (AC) in Sf9 cells and subsequent in vitro methods for assessing the activity of these cyclases. Membrane preparations derived from this overexpression system provide homogeneous sources of mammalian AC because AC that are endogenously expressed in Sf9 cells contribute low amounts of activity relative to the overexpressed enzyme. These approaches are applicable to all nine isoforms of mammalian membrane-bound AC isoforms.
D000262:Adenylyl Cyclases / Q000235:genetics* / Q000378:metabolism*; D000818:Animals; D002460:Cell Line; D007313:Insecta; D008322:Mammals; D008967:Molecular Biology / Q000379:methods*; D011994:Recombinant Proteins / Q000235:genetics* / Q000378:metabolism*; D015398:Signal Transduction
D011994:Recombinant Proteins; D000262:Adenylyl Cyclases