PXD062817-1

PXD062817 is an original dataset announced via ProteomeXchange.

Title	Structural Insights into Chemoresistance Mutants of BCL-2 and their Targeting by Stapled BAD BH3 Helices
Description	BCL-2 is a central regulator of apoptosis that inhibits cell death by sequestering pro-apoptotic BH3 alpha-helices within a hydrophobic surface groove. While venetoclax, a BH3-mimetic drug, has transformed the treatment of BCL-2–driven malignancies, its efficacy is increasingly limited by acquired resistance mutations that disrupt binding yet preserve anti-apoptotic function—a remarkable structural adaptation. Here, we employed hydrocarbon-stapled alpha-helices derived from the BAD BH3 motif as conformation-sensitive molecular probes to investigate this therapeutic challenge. The stapled peptides not only retain high-affinity binding to all BCL-2 variants, but also show enhanced potency to select venetoclax-resistant mutants. Structural analyses, including X-ray crystallography and hydrogen-deuterium exchange mass spectrometry (HDX MS), revealed the ability of stapled helices to restore native-like BH3 engagement by reverting the conformational consequences of resistance mutations. Notably, we identified a serendipitous interaction between the α3–α4 hairpin of BCL-2 and the hydrocarbon staple that compensates for altered groove conformation and contributes to mutant binding affinity. Together, these findings offer mechanistic insights into BCL-2 drug resistance and reveal a blueprint for designing next-generation inhibitors that overcome this clinically significant barrier to durable treatment responses.
HostingRepository	PRIDE
AnnounceDate	2025-09-30
AnnouncementXML	Submission_2025-09-30_12:35:48.394.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Thomas Wales
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	No PTMs are included in the dataset
Instrument	Synapt MS

Revision	Datetime	Status	ChangeLog Entry
0	2025-04-10 10:42:23	ID requested
⏵ 1	2025-09-30 12:35:48	announced

10.1038/S41467-025-63657-Y;

submitter keyword: BCL-2, venetoclax, BH3 inhibition, resistance mutation, hydrogen/deuterium exchange mass spectrometry, apoptotic regulation, BAD, HDX-MS, stapled peptide

Thomas E Wales
contact affiliation	Northeastern University
contact email	t.wales@northeastern.edu
lab head
Thomas Wales
contact affiliation	Northeastern University
contact email	t.wales@northeastern.edu
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2025/09/PXD062817

PRIDE project URI

• PRIDE
  1. PXD062817
     1. Label: PRIDE project
     2. Name: Structural Insights into Chemoresistance Mutants of BCL-2 and their Targeting by Stapled BAD BH3 Helices