PXD069511

PXD069511 is an original dataset announced via ProteomeXchange.

Title	Tryptoline Stereoprobe Elaboration Identifies Inhibitors of the GRPEL1-HSPA9 Chaperone Complex
Description	Activity-based protein profiling has identified hundreds of proteins from diverse classes that react site-specifically with stereo-defined electrophilic compounds (stereoprobes) in human cells. The structure-activity relationships underlying these stereoprobe-protein interactions, however, remain poorly understood. Here we show that the protein interaction landscape of tryptoline acrylamide stereoprobes can be profoundly altered by structural modifications distal to the acrylamide reactive group. Stereoprobe liganding events, many of which occurred at non-orthosteric sites, mostly evaded assignment by affinity prediction models (e.g., Boltz-2), which instead tended to redirect the ligands to orthosteric pockets (an outcome we refer to as “orthostery burnout”). We discovered that stereoprobes reacting with C124 in the nucleotide exchange factor GRPEL1 disrupt interactions with the mitochondrial Hsp70 chaperone (mortalin/HSPA9), leading to impairments in mitochondrial protein import and induction of mitophagy. Our results highlight tryptoline acrylamides as a versatile source of first-in-class covalent ligands targeting non-orthosteric sites on proteins, including tool compounds that perturb the mitochondrial HSP70 chaperone system.
HostingRepository	PRIDE
AnnounceDate	2025-10-22
AnnouncementXML	Submission_2025-10-21_16:24:35.453.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Rachel Hayward
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: NEWT:9606;
ModificationList	iodoacetamide derivatized residue
Instrument	Orbitrap Eclipse; Orbitrap Fusion

Revision	Datetime	Status	ChangeLog Entry
0	2025-10-15 23:15:11	ID requested
⏵ 1	2025-10-21 16:24:36	announced

10.1101/2025.10.20.683548;

submitter keyword: ABPP,chemical biology, stereoprobes

Benjamin F. Cravatt
contact affiliation	Department of Chemistry, Scripps Research, La Jolla, CA, USA
contact email	cravatt@scripps.edu
lab head
Rachel Hayward
contact affiliation	Scripps Research
contact email	rhayward@scripps.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/2025/10/PXD069511

PRIDE project URI

• PRIDE
  1. PXD069511
     1. Label: PRIDE project
     2. Name: Tryptoline Stereoprobe Elaboration Identifies Inhibitors of the GRPEL1-HSPA9 Chaperone Complex