PREreviews of “VHL ligand binding increases intracellular level of VHL” | PREreview

Skip to preprint details Skip to PREreviews

VHL ligand binding increases intracellular level of VHL

by Julianty Frost, Sonia Rocha, and Alessio Ciulli

Posted: April 12, 2021
Server: bioRxiv
DOI: 10.1101/2021.04.12.439487

Abstract

The von–Hippel Lindau (VHL) protein is a tumour suppressor protein frequently mutated in the VHL disease, which functions as substrate recognition subunit of a Cul2 E3 ubiquitin ligase (CRL2^VHL). CRL2^VHLplays an important role in oxygen sensing, by binding and targeting Hypoxia Inducible Factor-alpha subunits (HIF-alpha) for ubiquitination and degradation. VHL is also commonly hijacked by heterobifunctional degrader molecules known as proteolysis-targeting chimeras (PROTACs). In previous work we reported the structure-based design and functional characterisation of VHL inhibitors (VH032 and VH298) that induce the HIF response in cells. Here, we use unbiased quantitative mass spectrometry to identify the proteomic changes elicited by the VHL inhibitor and compare this to hypoxia or broad-spectrum prolyl-hydroxylase domain (PHD) enzyme inhibitor IOX2. Our results demonstrate the VHL inhibitor selectively activates the HIF response that is also present in the hypoxia- and IOX2-induced proteomic changes. Interestingly, VHL inhibitors were found to selectively upregulate a single protein, which is VHL itself. Our analysis revealed that this occurs via protein stabilisation of VHL isoforms and not via changes in transcript levels. Increased VHL levels upon VH298 treatment resulted in turn to reduced levels of HIF-1α protein. Our results demonstrate the high specificity of VHL inhibitors and suggest that use of these inhibitors would not produce overtly side effects due to prolonged HIF stabilisation. They also exemplify the concept that small-molecule binding induced protein stabilisation can increase protein levels inside cells.

Read the preprint

1 Rapid PREreview

by Milka Kostic

Where can I fill out a Rapid PREreview?

We’ve recently replaced Rapid PREreviews with Structured PREreviews.

You can write a Structured PREreview now.

We’ll be updating older Rapid PREreviews like these soon.

Aggregated Rapid PREreviews
Question	Yes	Unsure	N/A	No
Are the findings novel?	100%	0%	0%	0%
Are the results likely to lead to future research?	100%	0%	0%	0%
Is sufficient detail provided to allow reproduction of the study?	0%	100%	0%	0%
Are the methods and statistics appropriate for the analysis?	100%	0%	0%	0%
Are the principal conclusions supported by the data and analysis?	100%	0%	0%	0%
Does the manuscript discuss limitations?	100%	0%	0%	0%
Have the authors adequately discussed ethical concerns?	0%	0%	100%	0%
Does the manuscript include new data?	100%	0%	0%	0%
Are the data used in the manuscript available?	0%	0%	100%	0%
Is the code used in the manuscript available?	0%	0%	100%	0%
Would you recommend this manuscript to others?	100%	0%	0%	0%
Do you recommend this manuscript for peer review?	100%	0%	0%	0%

1 PREreview

Write a PREreview Request a PREreview

PREreview by Milka Kostic

Authored by Milka Kostic

GENERAL COMMENTS ON PREPRINT BY Frost, Rocha and Ciulli
In this preprint, Frost, Rocha and Ciulli report results of how cells respond to compounds previously described as inhibitors of von Hippel–Lindau (VHL) protein as measured by quantitative mass spectrometry (MS)-based proteomics. VHL is a …

Read the PREreview by Milka Kostic