干旱区生态环境知识资源中心(Arid): Development of a Sample-Preparation Workflow for Sulfopeptide Enrichment: From Target Analysis to Challenges in Shotgun Sulfoproteomics

Arid

DOI	10.1021/acs.analchem.0c01342
	Development of a Sample-Preparation Workflow for Sulfopeptide Enrichment: From Target Analysis to Challenges in Shotgun Sulfoproteomics
	Capriotti, Anna Laura 1; Cerrato, Andrea 1; Lagana, Aldo 1,2; Montone, Carmela Maria 1; Piovesana, Susy 1; Chiozzi, Riccardo Zenezini 3,4,5; Cavaliere, Chiara 1
通讯作者	Piovesana, Susy
来源期刊	ANALYTICAL CHEMISTRY
ISSN	0003-2700
EISSN	1520-6882
出版年	2020
卷号	92 期号:11 页码:7964-7971
英文摘要	Protein tyrosine O-sulfation is an important post-translational modification, as it has been correlated to inflammation, virus infection, and signal pathways. Nevertheless, methods for the characterization of protein sulfation by sulfopeptide enrichment are currently limited. In this Article, two standard compounds, representative of mono- and disulfated peptides, were used to compare the enrichment capabilities of five sorbent materials: two commercial weak anion-exchange mixed-mode sorbents (Strata X-AW and Oasis WAX) and three phosphopeptide enrichment materials based on affinity chromatography to either immobilized metals (IMAC) or metal oxides, i.e., Fe3+, TiO2, or Ti4+. The sulfopeptides were analyzed by ultrahigh-performance liquid chromatography (UHPLC) multiple-reaction monitoring analysis and were stable under all the tested experimental conditions. Recoveries of the enrichment step from spiked bovine serum albumin digests were >80% for the commercial Fe-IMAC kit and the Strata X-AW sorbent. Shotgun proteomics was used on the same samples to evaluate the selectivity, calculated as the number of coenriched peptides, and it was found to be better for the Fe-IMAC kit. Therefore, the Fe-IMAC protocol was embedded in a shotgun-proteomics workflow and applied to serum spiked with the sulfopeptides before protein dephosphorylation and digestion. The recovery of the entire analytical workflow was 20%, which was compatible with previous data on TiO2 phosphopeptide enrichment. Despite the potential, no sulfopeptide was confidently identified in serum digests by conventional shotgun proteomics, probably due to very low abundance of native sulfoproteins, poor ionization efficiency of sulfopeptides in the positive mode, and lack of unambiguous sulfopeptide identification by bioinformatics software. In this context, the use of negative-ionization mode with high-resolution mass spectrometry appeared promising to improve the sensibility and allow sulfopeptide identification in complex samples.
类型	Article
语种	英语
国家	Italy ; Netherlands
收录类别	SCI-E
WOS记录号	WOS:000538417400079
WOS关键词	MASS-SPECTROMETRIC DETECTION ; TYROSINE SULFATION ; ELECTRON-CAPTURE ; PEPTIDES ; IDENTIFICATION ; PROTEINS ; PLATFORM ; SITES ; PHOSPHORYLATION ; DISCRIMINATION
WOS类目	Chemistry, Analytical
WOS研究方向	Chemistry
资源类型	期刊论文
条目标识符	http://119.78.100.177/qdio/handle/2XILL650/319292
作者单位	1.Sapienza Univ Roma, Dept Chem, I-00185 Rome, Italy; 2.Univ Salento, CNR NANOTEC, Campus Ecotekne,Via Monteroni, I-73100 Lecce, Italy; 3.Univ Utrecht, Bijvoet Ctr Biomol Res, Biomol Mass Spectrometry & Prote, NL-3584 CH Utrecht, Netherlands; 4.Univ Utrecht, Utrecht Inst Pharmaceut Sci, NL-3584 CH Utrecht, Netherlands; 5.Netherlands Prote Ctr, NL-3584 CH Utrecht, Netherlands
推荐引用方式 GB/T 7714	Capriotti, Anna Laura,Cerrato, Andrea,Lagana, Aldo,et al. Development of a Sample-Preparation Workflow for Sulfopeptide Enrichment: From Target Analysis to Challenges in Shotgun Sulfoproteomics[J],2020,92(11):7964-7971.
APA	Capriotti, Anna Laura.,Cerrato, Andrea.,Lagana, Aldo.,Montone, Carmela Maria.,Piovesana, Susy.,...&Cavaliere, Chiara.(2020).Development of a Sample-Preparation Workflow for Sulfopeptide Enrichment: From Target Analysis to Challenges in Shotgun Sulfoproteomics.ANALYTICAL CHEMISTRY,92(11),7964-7971.
MLA	Capriotti, Anna Laura,et al."Development of a Sample-Preparation Workflow for Sulfopeptide Enrichment: From Target Analysis to Challenges in Shotgun Sulfoproteomics".ANALYTICAL CHEMISTRY 92.11(2020):7964-7971.