PXD025132

PXD025132 is an original dataset announced via ProteomeXchange.

Title	Simple Strategy of Integrating the Micro-centrifugation-based Extraction and 4-Hexylphenylazosulfonate for the Preparation of Plant Microsomal Proteins for Bottom-Up Proteomics
Description	The preparation of high-quality microsomal proteins (MPs) is of critical importance to high-throughput microsomal proteomics. The isolation of MPs for proteomic analysis has to date relied on Ultracentrifugation-based methods (UBMs), however, these methods are labor-intensive, time-consuming, and unaffordable to many laboratories. Here, we report a simple method developed by integrating the micro-centrifugation-based extraction (MME) method and 4-hexylphenylazosulfonate (Azo) surfactant for the isolation and solubilization of MPs from rice leaves. A mass spectrometry-based (MS) label-free proteomic approach was used for a comprehensive quantitative comparison of the extractability between the MME method and the plasma membrane extraction (PME) kit and of the ability to solubilize proteins between Azo and sodium dodecyl sulfate (SDS). Results showed that the MME method can not only effectively isolate MPs from rice leaves for a bottom-up proteomic analysis but also performs better than the PME kit in the enrichment of plasma membrane (PM) proteins. Additionally, Azo was found to effectively solubilize total proteins, including the MPs from rice leaves for bottom-up proteomics with performance comparable to that of SDS. Azo can be integrated with the MME approach as a simple and effective strategy to prepare MPs and could replace SDS for high-throughput proteomic studies. This is the first study revealing the applicability of the micro-centrifugation-based method and Azo in plant proteomics
HostingRepository	PRIDE
AnnounceDate	2021-09-29
AnnouncementXML	Submission_2021-09-28_18:34:07.954.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Sun Tae Kim
SpeciesList	scientific name: Oryza sativa Japonica Group; NCBI TaxID: 39947;
ModificationList	monohydroxylated residue; iodoacetamide derivatized residue
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2021-03-31 11:07:35	ID requested
⏵ 1	2021-09-28 18:34:08	announced

Nguyen TV, Gupta R, Annas D, Yoon J, Kim YJ, Lee GH, Jang JW, Park KH, Rakwal R, Jung KH, Min CW, Kim ST, An Integrated Approach for the Efficient Extraction and Solubilization of Rice Microsomal Membrane Proteins for High-Throughput Proteomics. Front Plant Sci, 12():723369(2021) [pubmed]

submitter keyword: Rice Leaves, Label-free Quantitative Proteomics, Microsomal Membrane Extraction, Plasma Membrane, Micro-centrifugation, Ultracentrifugation, Azo, SDS, MaxQuant, Perseus

Sun Tae Kim
contact affiliation	Department of Plant Bioscience, Pusan National University, Miryang 50463, South Korea
contact email	stki5505@gmail.com
lab head
Sun Tae Kim
contact affiliation	Pusan National University
contact email	stkim5505@gmail.com
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD025132
     1. Label: PRIDE project
     2. Name: Simple Strategy of Integrating the Micro-centrifugation-based Extraction and 4-Hexylphenylazosulfonate for the Preparation of Plant Microsomal Proteins for Bottom-Up Proteomics