Products/Services Used | Details | Operation |
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Custom Vector Construction> | A glycerol stock of transformed BL21(DE3) Rosetta™ cells with Bcl-2 encoded in a pET-15b vector (Novagen) was ordered from GenScript (Leiden, Netherlands) | Get A Quote |
B-cell lymphoma 2 (Bcl-2) proteins are the main regulators of mitochondrial apoptosis. Anti-apoptotic Bcl-2 proteins possess a hydrophobic tail-anchor enabling them to translocate to their target membrane and to shift into an active conformation where they inhibit pro-apoptotic Bcl-2 proteins to ensure cell survival. To address the unknown molecular basis of their cell-protecting functionality, we used intact human Bcl-2 protein natively residing at the mitochondrial outer membrane and applied neutron reflectometry and NMR spectroscopy. Here we show that the active full-length protein is entirely buried into its target membrane except for the regulatory flexible loop domain (FLD), which stretches into the aqueo... More