PXD018219 is an
original dataset announced via ProteomeXchange.
Dataset Summary
Title | Inter- and Intra-Molecular Interactions of The Arabidopsis Plasma Membrane Proton Pump Revealed Using a Mass Spectrometric-Cleavable Crosslinker |
Description | In plants and fungi, the plasma membrane proton pump (H+-ATPase) establishes an electrochemical gradient across the plasma membrane which serves as the driving force for the secondary transport of ions and nutrients across the cell membrane. This is an essential enzyme that functions in many important processes including stomatal movement, cell elongation, and cellular responses to stimuli from hormones, light, and other environmental conditions. Therefore, understanding how the activity of the H+-ATPase is regulated is important to understand how plants adapt to different growth conditions. The autoinhibitory effect of the C- terminal regulatory domain of H+-ATPase is well established and is thought to be mediated by interactions with the catalytic domains. Here, using the lysine reactive mass spectrometry cleavable crosslinker DSSO, we found that the C-terminal domain of the Arabidopsis H+- ATPase 2 (AHA2) crosslinked extensively with the actuator, nucleotide-binding, and phosphorylation domains, suggesting that the C-terminal domain regulates the catalytic cycle by modulating the relative positions of these domains. Interestingly, several C-terminal crosslinks occurred near a predicted proton binding site (Asp-684 in TM6), suggesting that the C-terminal domain may regulate proton efflux. Additionally, crosslinks between the C-terminal domain and other domains of AHA2 were detected in a monomeric protein resolved on SDS-PAGE, suggesting that intramolecular interactions may also be involved in the regulation of enzyme activity. Finally, we observed mixed-isotope crosslinking between unlabeled (14N-AHA2) and labeled (15N-AHA2) between the C-terminal domain and other domains of AHA2, supporting our model that oligomeric H+-ATPase may autoinhibit the neighboring monomer in a “head to tail” configuration. |
HostingRepository | PRIDE |
AnnounceDate | 2023-11-14 |
AnnouncementXML | Submission_2023-11-14_08:48:26.013.xml |
DigitalObjectIdentifier | |
ReviewLevel | Peer-reviewed dataset |
DatasetOrigin | Original dataset |
RepositorySupport | Unsupported dataset by repository |
PrimarySubmitter | Thao Thi Nguyen |
SpeciesList | scientific name: Arabidopsis thaliana (Mouse-ear cress); NCBI TaxID: 3702; scientific name: Saccharomyces cerevisiae (Baker's yeast); NCBI TaxID: 4932; |
ModificationList | monohydroxylated residue; iodoacetamide derivatized residue; deamidated residue |
Instrument | Orbitrap Fusion Lumos |
Dataset History
Revision | Datetime | Status | ChangeLog Entry |
0 | 2020-03-26 15:17:58 | ID requested | |
1 | 2023-06-01 11:49:54 | announced | |
⏵ 2 | 2023-11-14 08:48:29 | announced | 2023-11-14: Updated project metadata. |
Publication List
Nguyen TT, Blackburn MR, Sussman MR, Plasma Membrane Proton Pump Revealed Using a Mass Spectrometry Cleavable Cross-Linker. Biochemistry, 59(24):2210-2225(2020) [pubmed] |
Keyword List
submitter keyword: Arabidopsis, plasma membrane protein pump, metabolic labeling,DSSO, crosslinking |
Contact List
Michael R. Sussman |
contact affiliation | University of Wisconsin Madison, Department of Biochemistry |
contact email | msussman@wisc.edu |
lab head | |
Thao Thi Nguyen |
contact affiliation | University of Missouri-Columbia |
contact email | tnguyen24@wisc.edu |
dataset submitter | |
Full Dataset Link List
Dataset FTP location
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PRIDE project URI |
Repository Record List
[ + ]
[ - ]
- PRIDE
- PXD018219
- Label: PRIDE project
- Name: Inter- and Intra-Molecular Interactions of The Arabidopsis Plasma Membrane Proton Pump Revealed Using a Mass Spectrometric-Cleavable Crosslinker