PXD041372

PXD041372 is an original dataset announced via ProteomeXchange.

Title	Multisite phosphorylation by protein kinase A mediates modality-specific modulation of PIEZO2
Description	Protein kinase A is a downstream effector of many inflammatory mediators that induce pain hypersensitivity by increasing the mechanosensitivity of nociceptive sensory afferent. Here we examine the molecular mechanism underlying protein kinase-A dependent modulation of the mechanically-activated ion channel PIEZO2, which confers mechanosensitivity to many nociceptors. Using phosphorylation site prediction algorithms, we identified multiple putative and highly conserved PKA phosphorylation sites located on intracellular intrinsically disordered regions of PIEZO2. Site-directed mutagenesis and patch-clamp recordings showed that substitution of one or multiple putative PKA sites within a single intracellular domains does not alter PKA-induced PIEZO2 sensitization, whereas mutation of a combination of nine putative sites located on four different intracellular regions completely abolishes PKA-dependent PIEZO2 modulation, suggesting that the effect requires multisite phosphorylation. By demonstrating that PIEZO1 is not modulated by PKA, our data also reveals a previously unrecognized functional difference between PIEZO1 and PIEZO2. Moreover, by demonstrating that PKA only modulates PIEZO2 currents evoked by focal mechanical indentation of the cell, but not currents evoked by pressure-induced membrane stretch, we provide evidence suggesting that PIEZO2 is a polymodal mechanosensor that engages different protein domains for detecting different types of mechanical stimuli.
HostingRepository	PRIDE
AnnounceDate	2023-07-20
AnnouncementXML	Submission_2023-07-20_14:06:14.352.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	ThomasMair
SpeciesList	scientific name: Mus musculus (Mouse); NCBI TaxID: 10090;
ModificationList	phosphorylated residue
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2023-04-05 23:53:25	ID requested
⏵ 1	2023-07-20 14:06:15	announced

Schaefer I, Verkest C, Vespermann L, Mair T, Vo, ß H, Zeitzschel N, Lechner SG, PKA mediates modality-specific modulation of the mechanically gated ion channel PIEZO2. J Biol Chem, 299(6):104782(2023) [pubmed]

submitter keyword: LC-MS/MS, PIEZO2, Protein Kinase A, Proteomics, Phosphorylation

Stefan GLechner
contact affiliation	Department of Anesthesiology, University Medical Center Hamburg-Eppendorf, Martinistrasse 52, 20246, Hamburg, Germany.
contact email	s.lechner@uke.de
lab head
ThomasMair
contact affiliation	Section for Mass Spectrometry and Proteomics, University Medical Center Hamburg-Eppendorf
contact email	t.mair@uke.de
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD041372
     1. Label: PRIDE project
     2. Name: Multisite phosphorylation by protein kinase A mediates modality-specific modulation of PIEZO2