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Komagataella phaffii Erp41 is a protein disulfide isomerase with unprecedented disulfide bond catalyzing activity when coupled to glutathione

Palma, Arianna, Rettenbacher, Lukas A., Moilanen, Antti, Saaranen, Mirva, Gasser, Brigitte, Ruddock, Lloyd W. (2024) Komagataella phaffii Erp41 is a protein disulfide isomerase with unprecedented disulfide bond catalyzing activity when coupled to glutathione. The Journal of Biological Chemistry, . Article Number 105746. ISSN 0021-9258. E-ISSN 1083-351X. (doi:10.1016/j.jbc.2024.105746) (KAR id:105147)

Abstract

In the methylotrophic yeast Komagataella phaffii, we identified an ER-resident protein disulfide isomerase (PDI) family member, Erp41, with a peculiar combination of active site motifs. Like fungal ERp38, it has two thioredoxin-like domains which contain active site motifs (a and a'), followed by an alpha-helical ERp29c C-terminal domain (c domain). However, while the a domain has a typical PDI-like active site motif (CGHC), the a' domain instead has CGYC, a glutaredoxin-like motif which confers to the protein an exceptional affinity for GSH/GSSG. This combination of active site motifs has so far been unreported in PDI-family members. Homology searches revealed ERp41 is present in the genome of some plants, fungal parasites and a few non-conventional yeasts, among which are Komagataella spp. and Yarrowia lipolytica. These yeasts are both used for the production of secreted recombinant proteins. Here, we analyzed the activity of K. phaffii Erp41. We report that it is non-essential in K. phaffii, and that it can catalyze disulfide bond formation in partnership with the sulfhydryl oxidase Ero1 in vitro with higher turnover rates than the canonical PDI from K. phaffii, Pdi1, but slower activation times. We show how Erp41 has unusually fast glutathione-coupled oxidation activity and relate it to its unusual combination of active sites in its thioredoxin-like domains. We further describe how this determines its unusually efficient catalysis of dithiol oxidation in peptide and protein substrates.

Item Type: Article
DOI/Identification number: 10.1016/j.jbc.2024.105746
Uncontrolled keywords: glutathione, unfolded protein response (UPR), protein folding, yeast, endoplasmic reticulum (ER), Disulfide, enzyme catalysis, protein disulfide isomerase
Subjects: Q Science
Divisions: Divisions > Division of Natural Sciences > Biosciences
SWORD Depositor: JISC Publications Router
Depositing User: JISC Publications Router
Date Deposited: 28 Feb 2024 11:59 UTC
Last Modified: 29 Feb 2024 10:30 UTC
Resource URI: https://kar.kent.ac.uk/id/eprint/105147 (The current URI for this page, for reference purposes)

University of Kent Author Information

Rettenbacher, Lukas A..

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