PXD031833-1

PXD031833 is an original dataset announced via ProteomeXchange.

Title	TTYH family members form tetrameric complexes at the cell membrane
Description	The conserved Tweety homolog (TTYH) family consists of three paralogs in vertebrates, displaying a ubiquitous expression pattern. Although considered as ion channels for almost two decades, recent structural and functional analyses refuted this role. Intriguingly, while all paralogs shared a dimeric stoichiometry following detergent solubilization, their structures revealed divergence in their relative subunit orientation. Here, we determined the stoichiometry of intact mouse TTYH (mTTYH) complexes in cells. Using cross-linking and single-molecule fluorescence microscopy, we demonstrate that mTTYH1 and mTTYH3 form tetramers at the plasma membrane, stabilized by interactions between their extracellular domains. Using blue-native PAGE, fluorescence-detection size-exclusion chromatography and hydrogen-deuterium exchange mass spectrometry (HDX-MS), we reveal that detergent solubilization results in tetramers destabilization, leading to their dissolution into dimers. Moreover, HDX-MS demonstrates that the extracellular domains are stabilized in the context of the tetrameric mTTYH complex. Together, our results expose the innate tetrameric organization of TTYH complexes at the cell membrane. Future structural analyses of these assemblies in native membranes are required to illuminate their long-sought cellular function.
HostingRepository	PRIDE
AnnounceDate	2022-08-31
AnnouncementXML	Submission_2022-08-31_08:02:39.992.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Petr Man
SpeciesList	scientific name: Mus musculus (Mouse); NCBI TaxID: 10090;
ModificationList	N-glycosylated residue; acetylated residue; glycosylated residue
Instrument	timsTOF Pro; Bruker Daltonics solarix series

Revision	Datetime	Status	ChangeLog Entry
0	2022-02-22 13:07:07	ID requested
⏵ 1	2022-08-31 08:02:40	announced
2	2023-11-14 08:55:50	announced	2023-11-14: Updated project metadata.

10.1038/S42003-022-03862-3;

submitter keyword: Tweety homolog family, TTYH, hydrogen/deuterium exchange, dimer, tetramer

Petr Man
contact affiliation	Petr Man, PhD Laboratory of Structural Biology and Cell Signaling BioCeV - Institute of Microbiology of the Czech Acad Sci Prumyslova 595 252 50 Vestec Czech Republic
contact email	pman@biomed.cas.cz
lab head
Petr Man
contact affiliation	Institute of Microbiology of the CAS
contact email	pman@biomed.cas.cz
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD031833
     1. Label: PRIDE project
     2. Name: TTYH family members form tetrameric complexes at the cell membrane