Version 1
: Received: 15 August 2024 / Approved: 16 August 2024 / Online: 16 August 2024 (08:28:11 CEST)
How to cite:
Hoang, M. N.; Peterbauer, C. Roles of Mature Domain Targeting Signals for Protein Translocation and Secretion in Lactococcus lactis. Preprints2024, 2024081219. https://doi.org/10.20944/preprints202408.1219.v1
Hoang, M. N.; Peterbauer, C. Roles of Mature Domain Targeting Signals for Protein Translocation and Secretion in Lactococcus lactis. Preprints 2024, 2024081219. https://doi.org/10.20944/preprints202408.1219.v1
Hoang, M. N.; Peterbauer, C. Roles of Mature Domain Targeting Signals for Protein Translocation and Secretion in Lactococcus lactis. Preprints2024, 2024081219. https://doi.org/10.20944/preprints202408.1219.v1
APA Style
Hoang, M. N., & Peterbauer, C. (2024). Roles of Mature Domain Targeting Signals for Protein Translocation and Secretion in <em>Lactococcus lactis</em>. Preprints. https://doi.org/10.20944/preprints202408.1219.v1
Chicago/Turabian Style
Hoang, M. N. and Clemens Peterbauer. 2024 "Roles of Mature Domain Targeting Signals for Protein Translocation and Secretion in <em>Lactococcus lactis</em>" Preprints. https://doi.org/10.20944/preprints202408.1219.v1
Abstract
Lactococcus lactis is a potential bacterial cell factory to develop delivery systems for vaccines and therapeutic proteins. Large progress has been made in applications using engineered L. lactis against, e.g., inflammatory bowel disease and cervical cancer, but improvement of secretion and cell anchoring efficacy is still desirable. A double-labelling method based on biarsenical hairpin binding and nickel-polyhistidine affinity was used for visualization of protein trafficking and quantification of targeted proteins on the cell surface and in the cytoplasm. To investigate the importance of mature domain targeting signals (MTSs), we generated truncated constructs encoding 126, 66, and 26 amino acid residues from the N-terminus of the basic membrane protein A (BmpA) and fused those with the gene for the human papillomavirus serotype 16 (HPV16) E7 oncoprotein. Overexpression of fusion proteins was observed to come at a cost of cell proliferation. L. lactis cells produced and displayed the shortest fusion protein only with difficulty, suggesting that the entire absence of a homologous sequence containing MTSs significantly impedes export and surface anchoring of fusion proteins. With 40 amino acids following the signal peptide and containing one MTS, effective translocation was possible. Mutations of MTSs towards increased hydrophobicity resulted in an increase of secreted and surface-displayed fusion protein, suggesting the potential to design rationally improved constructs.
Biology and Life Sciences, Biochemistry and Molecular Biology
Copyright:
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
