PXD032052-1

PXD032052 is an original dataset announced via ProteomeXchange.

Title	Identification of PPT1 substrates highlights roles of depalmitoylation in disulfide bond formation and synaptic function
Description	Loss-of-function mutations in the depalmitoylating enzyme palmitoyl protein thioesterase 1 (PPT1) cause Neuronal Ceroid Lipofuscinosis (NCL), a devastating neurodegenerative disease. The substrates of PPT1 are largely undescribed, posing a limitation on molecular dissection of disease mechanisms and therapeutic development. Here, we provide a resource identifying >100 novel PPT1 substrates. We utilized Acyl Resin-Assisted Capture and mass spectrometry to identify proteins with increased in vivo palmitoylation in PPT1 knockout mouse brains. We then validated putative substrates through direct depalmitoylation with recombinant PPT1. This stringent screen elucidated diverse PPT1 substrates at the synapse, including channels and transporters, G-protein-associated molecules, endo/exocytic components, synaptic adhesion molecules, and mitochondrial proteins. Cysteine depalmitoylation sites in transmembrane PPT1 substrates frequently participate in disulfide bonds in the mature protein. We confirmed that depalmitoylation plays a role in disulfide bond formation in a tertiary screen analyzing post-translational modifications. Collectively, these data highlight the role of PPT1 in mediating synapse functions, implicate molecular pathways in the etiology of NCL and other neurodegenerative diseases, and advance our basic understanding of the purpose of depalmitoylation.
HostingRepository	PRIDE
AnnounceDate	2022-03-07
AnnouncementXML	Submission_2022-03-06_20:19:06.789.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	TuKiet Lam
SpeciesList	scientific name: Mus musculus (Mouse); NCBI TaxID: 10090;
ModificationList	phosphorylated residue; monohydroxylated residue; acetylated residue; iodoacetamide derivatized residue; deamidated residue
Instrument	Q Exactive Plus

Revision	Datetime	Status	ChangeLog Entry
0	2022-03-03 12:23:45	ID requested
⏵ 1	2022-03-06 20:19:07	announced
2	2022-03-23 14:34:37	announced	2022-03-23: Updated project metadata.

Dataset with its publication pending

submitter keyword: palmitoylation, synapse, palmitoyl protein thioesterase 1, neuronal ceroid lipofuscinosis, neurodegeneration, synaptic adhesion molecules, synaptic vesicle endocytosis, ZDHHC, disulfide bond, Na+/K+ transporting ATPase

TuKiet Lam
contact affiliation	Yale University (Keck MS & Proteomics Resource)
contact email	tukiet.lam@yale.edu
lab head
TuKiet Lam
contact affiliation	Yale University
contact email	tukiet.lam@yale.edu
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD032052
     1. Label: PRIDE project
     2. Name: Identification of PPT1 substrates highlights roles of depalmitoylation in disulfide bond formation and synaptic function