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

Sandhu, Hena; Hedman, Rickard; Cymer, Florian; Kudva, Renuka; Ismail, Nurzian; Heijne, Gunnar von

doi:10.1016/j.jmb.2021.167047

Cited by 9 publications

(9 citation statements)

References 54 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…An interesting method to study in vivo insertion and co-translational folding of membrane proteins is the application of translational arrest peptides to measure forces acting on a nascent protein during membrane insertion ( Ismail et al, 2012 ; Sandhu et al, 2021 ). In this approach, the arrest peptide binds to the ribosomal tunnel and induces ribosomal stalling at a specific amino acid.…”

Section: Signal Peptidesmentioning

confidence: 99%

Bacterial Signal Peptides- Navigating the Journey of Proteins

Kaushik

Dalbey

2022

Front. Physiol.

View full text Add to dashboard Cite

In 1971, Blobel proposed the first statement of the Signal Hypothesis which suggested that proteins have amino-terminal sequences that dictate their export and localization in the cell. A cytosolic binding factor was predicted, and later the protein conducting channel was discovered that was proposed in 1975 to align with the large ribosomal tunnel. The 1975 Signal Hypothesis also predicted that proteins targeted to different intracellular membranes would possess distinct signals and integral membrane proteins contained uncleaved signal sequences which initiate translocation of the polypeptide chain. This review summarizes the central role that the signal peptides play as address codes for proteins, their decisive role as targeting factors for delivery to the membrane and their function to activate the translocation machinery for export and membrane protein insertion. After shedding light on the navigation of proteins, the importance of removal of signal peptide and their degradation are addressed. Furthermore, the emerging work on signal peptidases as novel targets for antibiotic development is described.

show abstract

Section: Signal Peptidesmentioning

confidence: 99%

Bacterial Signal Peptides- Navigating the Journey of Proteins

Kaushik

Dalbey

2022

Front. Physiol.

View full text Add to dashboard Cite

show abstract

“…The AP-induced stalling can be overcome by pulling forces acting on the nascent chain [14,15], with different APs being sensitive to different force levels [16]. Such pulling forces can be generated by, e.g., cotranslational protein folding in the cytoplasm as well as in the periplasm, or cotranslational insertion of transmembrane segments into the membrane [9,17,18]. Therefore, APs can be conveniently used as force sensors to study cotranslational events in vitro and in vivo.…”

Section: Force Profile Analysismentioning

confidence: 99%

Upstream charged and hydrophobic residues impact the timing of membrane insertion of transmembrane helices

et al. 2022

View full text Add to dashboard Cite

During SecYEG‐mediated cotranslational insertion of membrane proteins, transmembrane helices (TMHs) first make contact with the membrane when their N‐terminal end is ~ 45 residues away from the peptidyl transferase centre. However, we recently uncovered instances where the first contact is delayed by up to ~ 10 residues. Here, we recapitulate these effects using a model TMH fused to two short segments from the Escherichia coli inner membrane protein BtuC: a positively charged loop and a re‐entrant loop. We show that the critical residues are two Arg residues in the positively charged loop and four hydrophobic residues in the re‐entrant loop. Thus, both electrostatic and hydrophobic interactions involving sequence elements that are not part of a TMH can impact the way the latter behaves during membrane insertion.

show abstract

“…The translational arrest can be overcome if a strong enough pulling force is exerted on the AP, essentially pulling it out of its binding site in the exit tunnel (12)(13)(14)(15)(16). APs can be employed as sensitive 'molecular force sensors' to report on various cotranslational events such as protein folding (17,18), protein translocation (19,20), and membrane protein integration (7,13).…”

Section: Introductionmentioning

confidence: 99%

Cotranslational folding and assembly of the dimeric E. coli inner membrane protein EmrE

Mermans

Nicolaus

Fleisch

et al. 2022

Preprint

Self Cite

View full text Add to dashboard Cite

In recent years, it has become clear that many homo- and heterodimeric cytoplasmic proteins in both prokaryotic and eukaryotic cells start to dimerize cotranslationally, i.e., while at least one of the two chains is still attached to the ribosome. Whether this is possible also for integral membrane proteins is unknown, however. Here, we apply Force Profile Analysis (FPA) - a method where a translational arrest peptide (AP) engineered into the polypeptide chain is used to detect force generated on the nascent chain during membrane insertion - to demonstrate cotranslational interactions between a fully membrane-inserted monomer and a nascent, ribosome-tethered monomer of the E. coli inner membrane protein EmrE. Similar cotranslational interactions are also seen when the two monomers are fused into a single polypeptide. Further, we uncover an apparent intrachain interaction between E14 in TMH1 and S64 in TMH3 that forms at a precise nascent chain length during cotranslational membrane insertion of an EmrE monomer. Like soluble proteins, inner membrane proteins can thus both start to fold and start to dimerize during the cotranslational membrane-insertion process.

show abstract

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

Cited by 9 publications

References 54 publications

Bacterial Signal Peptides- Navigating the Journey of Proteins

Bacterial Signal Peptides- Navigating the Journey of Proteins

Upstream charged and hydrophobic residues impact the timing of membrane insertion of transmembrane helices

Cotranslational folding and assembly of the dimeric E. coli inner membrane protein EmrE

Contact Info

Product

Resources

About