Destabilization and Degradation of a Disease-Linked PGM1 Protein Variant

Frederik Gouliaev, Nicolas Jonsson, Sarah Gersing, Michael Lisby, Kresten Lindorff-Larsen*, Rasmus Hartmann-Petersen

*Corresponding author af dette arbejde

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

Abstract

PGM1-linked congenital disorder of glycosylation (PGM1-CDG) is an autosomal recessive disease characterized by several phenotypes, some of which are life-threatening. Research focusing on the disease-related variants of the α-D-phosphoglucomutase 1 (PGM1) protein has shown that several are insoluble in vitro and expressed at low levels in patient fibroblasts. Due to these observations, we hypothesized that some disease-linked PGM1 protein variants are structurally destabilized and subject to protein quality control (PQC) and rapid intracellular degradation. Employing yeast-based assays, we show that a disease-associated human variant, PGM1 L516P, is insoluble, inactive, and highly susceptible to ubiquitylation and rapid degradation by the proteasome. In addition, we show that PGM1 L516P forms aggregates in S. cerevisiae and that both the aggregation pattern and the abundance of PGM1 L516P are chaperone-dependent. Finally, using computational methods, we perform saturation mutagenesis to assess the impact of all possible single residue substitutions in the PGM1 protein. These analyses identify numerous missense variants with predicted detrimental effects on protein function and stability. We suggest that many disease-linked PGM1 variants are subject to PQC-linked degradation and that our in silico site-saturated data set may assist in the mechanistic interpretation of PGM1 variants.
OriginalsprogEngelsk
TidsskriftBiochemistry
Vol/bind63
Udgave nummer11
Sider (fra-til)1423–1433
Antal sider11
ISSN0006-2960
DOI
StatusUdgivet - 2024

Bibliografisk note

Funding Information:
The present work was funded by the Novo Nordisk Foundation ( https://novonordiskfonden.dk ) challenge program PRISM (to K.L.-L. and R.H.-P.), and the Danish Council for Independent Research (Det Frie Forskningsr\u00E5d) ( https://dff.dk ) 10.46540/2032\u201300007B (to R.H.P.). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Publisher Copyright:
© 2024 American Chemical Society.

Citationsformater