PXD014137

PXD014137 is an original dataset announced via ProteomeXchange.

Title	Rapid single-step affinity purification of HA-tagged mitochondria from Arabidopsis thaliana
Description	Photosynthesis in plant cells would not be possible without the supportive role of mitochondria. However, isolating mitochondria for physiological and biochemical analyses from plant cells is a lengthy and tedious process. Established isolation protocols require multiple centrifugation steps and substantial amounts of starting material. To overcome these limitations, we tagged mitochondria in plant cells with a triple haemagglutinin-tag for rapid purification by a single affinity purification step. This protocol yields highly purified mitochondria from 1 g of Arabidopsis seedlings that are suitable for enzyme activity measurements and delivers sufficient amounts of mitochondrial proteins for deep proteomic profiling. We demonstrate that the method can be applied to proteomic analysis of an Arabidopsis mutant deficient in the mitochondrial glutamate transporter À bout de souffle (BOU) and we identify 27 differentially expressed proteins in the bou mutant. Our work also sets the stage for the development of advanced isolation protocols for mitochondria from distinct cell types.
HostingRepository	PRIDE
AnnounceDate	2023-11-14
AnnouncementXML	Submission_2023-11-14_09:14:30.385.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Nina Overbeck
SpeciesList	scientific name: Arabidopsis thaliana (Mouse-ear cress); NCBI TaxID: 3702;
ModificationList	monohydroxylated residue; iodoacetamide derivatized residue
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2019-06-05 02:26:14	ID requested
1	2023-11-14 00:16:04	announced
⏵ 2	2023-11-14 09:14:31	announced	2023-11-14: Updated project metadata.

Kuhnert F, Stefanski A, Overbeck N, Drews L, Reichert AS, St, ü, hler K, Weber APM, Rapid Single-Step Affinity Purification of HA-Tagged Plant Mitochondria. Plant Physiol, 182(2):692-706(2020) [pubmed]

curator keyword: Biological

submitter keyword: Arabidopsis,Mitochondria, LC-MS/MS

Kai Stuehler
contact affiliation	Institute for Molecular Medicine I, Medical Faculty, Heinrich-Heine-University Duesseldorf, Universitaetsstrasse 1, 40225 Duesseldorf, Germany
contact email	kai.stuehler@hhu.de
lab head
Nina Overbeck
contact affiliation	BMFZ/MPL
contact email	nina.overbeck@hhu.de
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2023/11/PXD014137

PRIDE project URI

• PRIDE
  1. PXD014137
     1. Label: PRIDE project
     2. Name: Rapid single-step affinity purification of HA-tagged mitochondria from Arabidopsis thaliana