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Proteins, Expression, Isolation and Analysis | Microsome aliquots were boiled for 5 min in 2× SDS buffer immediately after thawing. 0.5–1 units of microsomes were analyzed by SDS-PAGE and immunoblotting using anti-FLAG antibody (Genscript). | Get A Quote |
Molecular recognition features (MoRFs) provide interaction motifs in intrinsically disordered protein regions to mediate diverse cellular functions. Here we report that a MoRF element, located in the disordered linker domain of the mammalian signal recognition particle (SRP) receptor and conserved among eukaryotes, plays an essential role in sensing the ribosome during cotranslational protein targeting to the endoplasmic reticulum. Loss of the MoRF in the SRP receptor (SR) largely abolishes the ability of the ribosome to activate SRP-SR assembly and impairs cotranslational protein targeting. These results demonstrate a novel role for MoRF elements and provide a mechanism for the ribosome-induced activation of t... More