Role of the signal sequence in proteorhodopsin biogenesis in E. coli

Soto-Rodríguez, Jessica; Baneyx, François

doi:10.1002/bit.26878

Title: Role of the signal sequence in proteorhodopsin biogenesis in E. coli

Journal Article · 25 November 2018 · Biotechnology and Bioengineering

DOI: https://doi.org/10.1002/bit.26878 · OSTI ID:1611423

Soto-Rodríguez, Jessica ^[1];

^[2]

Univ. of Washington, Seattle, WA (United States); DOE/OSTI
Univ. of Washington, Seattle, WA (United States)

Blue-absorbing proteorhodopsin (BPR) from marine bacteria is a retinal-bound, light-activated, outwards proton transporter containing seven α-helical transmembrane segments (TMS). It is synthesized as a precursor species (pre-BPR) with a predicted N-terminal signal sequence that is cleaved to yield the mature protein. While optimizing the production of BPR in Escherichia coli to facilitate the construction of bioprotonic devices, we observed significant pre-BPR accumulation in the inner membrane and explored signal sequence requirements and export pathway. Here, we report that BPR does not rely on the Sec pathway for inner membrane integration, and that although it greatly enhances yields, its signal sequence is not necessary to obtain a functional product. We further show that an unprocessable version of pre-BPR obtained by mutagenesis of the signal peptidase I site exhibits all functional attributes of the wild-type protein and has the advantage of being produced at higher levels. Our results are consistent with the BPR signal sequence being recognized by the signal recognition particle (SRP; a protein that orchestrates the cotranslational biogenesis of inner membrane proteins) and serving as a beneficial “pro” domain rather than a traditional secretory peptide.

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Research Organization:: Univ. of Washington, Seattle, WA (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Basic Energy Sciences (BES)

Grant/Contract Number:: SC0010441

OSTI ID:: 1611423

Journal Information:: Biotechnology and Bioengineering, Journal Name: Biotechnology and Bioengineering Journal Issue: 4 Vol. 116; ISSN 0006-3592

Publisher:: WileyCopyright Statement

Country of Publication:: United States

Language:: English

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