PXD008226

PXD008226 is an original dataset announced via ProteomeXchange.

Title	An inter-subunit protein-protein interface that stabilizes the specific activity and oligomerization of the AAA+ chaperone Reptin
Description	Reptin forms multiple inter-subunit protein-protein interaction contacts. Rate-limiting motifs in ligand-dependent oligomer assembly are not defined. We set up a Reptin-self peptide scan assay to identify functional protein-protein interfaces that dominate in Reptin self assembly. The most dominant self-peptide binding mapped to the inter-subunit “rim” of the Reptin oligomer that is formed a tyrosine finger binding into an adjacent hydrophobic region in an adjacent subunit. Reptin binding to the tyrosine finger peptide motif is suppressed by Liddean or ADP suggesting that oligomer formation excludes the self-peptide from binding. HDX-mass spectrometry demonstrated that ATP suppressed deuteration at the dimer interface in both wt and Y340A Reptin. However, the Y340A mutation attenuated deuterium suppression of Reptin within the tyrosine finger in the presence of ligand. The tyrosine finger of Reptin interacts with a more shallow pocket in Pontin. Gel filtration demonstrated that the Y340A Reptin mutant does not form ADP-induced oligomers compared to the D299N mutant or wt- Reptin. The Y340A mutation shows increased AGR2 binding but decreased Pontin or p53 binding. These data indicate that the Y340 tyrosine “finger” plays an important role in stabilizing the Reptin oligomer in the presence of ligand. We propose that the Reptin interactome is regulated by ligand-dependent conversion between monomeric and oligomeric forms that is regulated by a divergent inter-subunit protein-protein interaction motif.
HostingRepository	PRIDE
AnnounceDate	2019-04-23
AnnouncementXML	Submission_2019-04-23_05:22:22.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Dominika Coufalová
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	No PTMs are included in the dataset
Instrument	LTQ Orbitrap Elite

Revision	Datetime	Status	ChangeLog Entry
0	2017-11-16 02:16:01	ID requested
⏵ 1	2019-04-23 05:22:23	announced

Coufalova D, Remnant L, Hernychova L, Muller P, Healy A, Kannan S, Westwood N, Verma CS, Vojtesek B, Hupp TR, Houston DR, An inter-subunit protein-peptide interface that stabilizes the specific activity and oligomerization of the AAA+ chaperone Reptin. J Proteomics, 199():89-101(2019) [pubmed]

curator keyword: Biological

submitter keyword: Reptin, oligomer assembly, protein-protein interaction, HDX mass spectrometry

Ted R. Hupp
contact affiliation	Edinburgh Cancer Research Centre, Cell Signalling Unit, University of Edinburgh
contact email	ted.hupp@ed.ac.uk
lab head
Dominika Coufalová
contact affiliation	d RECAMO, Masaryk Memorial Cancer Institute
contact email	coufalovadominika@gmail.com
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD008226
     1. Label: PRIDE project
     2. Name: An inter-subunit protein-protein interface that stabilizes the specific activity and oligomerization of the AAA+ chaperone Reptin