Membrane Targeting and Insertion of the C-Tail Protein SciP

•C-tailed protein SciP has a signal recognition particle-dependent targeting information at its N terminus.•The first 27 residues of SciP guide a green fluorescent protein fusion protein to the membrane.•Extension of the SciP C-tail by 10 residues involves YidC and Sec for insertion.

C-tailed membrane proteins insert into the bilayer post-translationally because the hydrophobic anchor segment leaves the ribosome at the end of translation. Nevertheless, we find here evidence that the targeting of SciP to the membrane of Escherichia coli occurs co-translationally since signal elements in the N-terminal part of the SciP protein sequence are present. Two short hydrophobic sequences were identified that targeted a green fluorescent protein-SciP fusion protein to the membrane involving the signal recognition particle. After targeting, the membrane insertion of SciP is catalyzed by YidC independent of the SecYEG translocase. However, when the C-terminal tail of SciP was extended to 21 aa residues, we found that SecYEG becomes involved and makes its membrane insertion more efficient.

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