Cotranslational Translocation and Folding of a Periplasmic Protein Domain in Escherichia coli

Sandhu, HenaStockholms universitet,Institutionen för biokemi och biofysik (author)

In Gram-negative bacteria, periplasmic domains in inner membrane proteins are cotranslationally translocated across the inner membrane through the SecYEG translocon. To what degree such domains also start to fold cotranslationally is generally difficult to determine using currently available methods. Here, we apply Force Profile Analysis (FPA) - a method where a translational arrest peptide is used to detect folding-induced forces acting on the nascent polypeptide - to follow the cotranslational translocation and folding of the large periplasmic domain of the E. coli inner membrane protease LepB in vivo. Membrane insertion of LepB's two N-terminal transmembrane helices is initiated when their respective N-terminal ends reach 45-50 residues away from the peptidyl transferase center (PTC) in the ribosome. The main folding transition in the periplasmic domain involves all but the similar to 15 most C-terminal residues of the protein and happens when the C-terminal end of the folded part is similar to 70 residues away from the PTC; a smaller putative folding intermediate is also detected. This implies that wildtype LepB folds post-translationally in vivo, and shows that FPA can be used to study both co- and post-translational protein folding in the periplasm.

Hedman, RickardStockholms universitet,Institutionen för biokemi och biofysik(Swepub:su)rihe4867 (author)
Cymer, FlorianStockholms universitet,Institutionen för biokemi och biofysik(Swepub:su)fcyme (author)
Kudva, RenukaStockholms universitet,Institutionen för biokemi och biofysik,Science for Life Laboratory (SciLifeLab)(Swepub:su)rkudv (author)
Ismail, NurzianStockholms universitet,Institutionen för biokemi och biofysik(Swepub:su)nisma (author)
von Heijne, GunnarStockholms universitet,Institutionen för biokemi och biofysik,Science for Life Laboratory (SciLifeLab)(Swepub:su)heijn (author)
Stockholms universitetInstitutionen för biokemi och biofysik (creator_code:org_t)

By the author/editor: Sandhu, Hena; Hedman, Rickard; Cymer, Florian; Kudva, Renuka; Ismail, Nurzian; von Heijne, Gunn ...

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