University of Oulu

Palma, A., Rettenbacher, L.A., Moilanen, A. et al. Biochemical analysis of Komagataella phaffii oxidative folding proposes novel regulatory mechanisms of disulfide bond formation in yeast. Sci Rep 13, 14298 (2023).

Biochemical analysis of Komagataella phaffii oxidative folding proposes novel regulatory mechanisms of disulfide bond formation in yeast

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Author: Palma, Arianna1,2; Rettenbacher, Lukas A.3,4; Moilanen, Antti4;
Organizations: 1Department of Biotechnology, University of Natural Resources and Life Sciences, Vienna, Austria
2Austrian Centre of Industrial Biotechnology, Vienna, Austria
3School of Biosciences, University of Kent, Canterbury, UK
4Faculty of Biochemistry and Molecular Medicine, University of Oulu, Oulu, Finland
Format: article
Version: published version
Access: open
Online Access: PDF Full Text (PDF, 2.8 MB)
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Language: English
Published: Springer Nature, 2023
Publish Date: 2023-10-09


Oxidative protein folding in the endoplasmic reticulum (ER) is driven mainly by protein disulfide isomerase PDI and oxidoreductin Ero1. Their activity is tightly regulated and interconnected with the unfolded protein response (UPR). The mechanisms of disulfide bond formation have mainly been studied in human or in the yeast Saccharomyces cerevisiae. Here we analyze the kinetics of disulfide bond formation in the non-conventional yeast Komagataella phaffii, a common host for the production of recombinant secretory proteins. Surprisingly, we found significant differences with both the human and S. cerevisiae systems. Specifically, we report an inactive disulfide linked complex formed by K. phaffii Ero1 and Pdi1, similarly to the human orthologs, but not described in yeast before. Furthermore, we show how the interaction between K. phaffii Pdi1 and Ero1 is unaffected by the introduction of unfolded substrate into the system. This is drastically opposed to the previously observed behavior of the human pathway, suggesting a different regulation of the UPR and/or possibly different interaction mechanics between K. phaffii Pdi1 and Ero1.

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Series: Scientific reports
ISSN: 2045-2322
ISSN-E: 2045-2322
ISSN-L: 2045-2322
Volume: 13
Article number: 14298
DOI: 10.1038/s41598-023-41375-z
Type of Publication: A1 Journal article – refereed
Field of Science: 1182 Biochemistry, cell and molecular biology
Funding: This work was funded by the Marie Skłodowska-Curie Actions Innovative Training Network of the European Union’s Horizon 2020 Program under grant agreement no. 813979 (SECRETERS). Further support was obtained by the Austrian Federal Ministry of Digital and Economic Affairs (BMDW), the Austrian Federal Ministry of Traffic, Innovation and Technology (bmvit), the Styrian Business Promotion Agency SFG, the Standortagentur Tirol, the Government of Lower Austria, the Business Agency Vienna and BOKU through the COMET Funding Program managed by the Austrian Research Promotion Agency FFG.
EU Grant Number: (813979) Secreters - A new generation of microbial expression hosts and tools for the production of biotherapeutics and high-value enzymes
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