PXD061523

PXD061523 is an original dataset announced via ProteomeXchange.

Title	Protein Complexes Using Dual-Tagged Protein Complex Isolation (DPCI) in Plant Systems
Description	Protein complexes are crucial for cellular processes, but identifying them is challenging due to mass spectrometry noise. Here, we developed a new method, Dual-tagged protein complex isolation (DPCI), to address this. In this method, the bait protein was fused with Strep tag II and Flag tag, then transformed into plants using a highly efficient transient transformation technique for optimal expression. The plants were cross-linked with formaldehyde to stabilize protein interactions. Proteins were extracted and incubated with Flag antibody, secondary antibody, and Strep-Tactin beads. The protein complex was purified using Strep-Tactin beads, and the bait protein was simultaneously labeled with Flag and secondary antibodies. The isolated protein complexes were subjected to agarose gel electrophoresis, and the gel was soaked in ECL buffer to visualize bait protein complexes. Gel pieces were cut out for recovery and LC-MS/MS. This method successfully isolated the protein complex of the BpEBP1 transcription factor from Betula platyphylla, identifying 53 proteins via LC-MS/MS. Further analysis, including Y2H assays, confirmed 19 interactions with 78.56% accuracy, elucidating the members of the BpEBP1 protein complex and its interaction network. This method can be used in any plant transformable by Agrobacterium, regardless of a stable genetic transformation system, offering broad applications.
HostingRepository	iProX
AnnounceDate	2025-03-06
AnnouncementXML	Submission_2025-03-05_22:45:34.062.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Shilin Sun
SpeciesList	scientific name: Betula platyphylla; NCBI TaxID: 78630;
ModificationList	No PTMs are included in the dataset
Instrument	Q Exactive HF

Revision	Datetime	Status	ChangeLog Entry
0	2025-03-05 22:45:23	ID requested
⏵ 1	2025-03-05 22:45:34	announced

Dataset with its publication pending

submitter keyword: Protein complexes, Transient transformation, Protein interaction network, Plant, Mass spectrometry, Protein identification

Yucheng Wang
contact affiliation	Shenyang Agricultural University
contact email	wangyucheng@ms.xjb.ac.cn
lab head
Shilin Sun
contact affiliation	Shenyang Agricultural University
contact email	18804056008@163.com
dataset submitter

iProX dataset URI