PMID- 32380835
OWN - NLM
STAT- MEDLINE
DCOM- 20210212
LR  - 20210212
IS  - 1520-6882 (Electronic)
IS  - 0003-2700 (Linking)
VI  - 92
IP  - 11
DP  - 2020 Jun 2
TI  - Development of a Sample-Preparation Workflow for Sulfopeptide Enrichment: From 
      Target Analysis to Challenges in Shotgun Sulfoproteomics.
PG  - 7964-7971
LID - 10.1021/acs.analchem.0c01342 [doi]
AB  - 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., Fe(3+), TiO(2), or Ti(4+). 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 
      TiO(2) 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.
FAU - Capriotti, Anna Laura
AU  - Capriotti AL
AUID- ORCID: 0000-0003-1017-9625
AD  - Department of Chemistry, Sapienza Universita di Roma, Piazzale Aldo Moro 5, 00185 
      Rome, Italy.
FAU - Cerrato, Andrea
AU  - Cerrato A
AD  - Department of Chemistry, Sapienza Universita di Roma, Piazzale Aldo Moro 5, 00185 
      Rome, Italy.
FAU - Lagana, Aldo
AU  - Lagana A
AD  - Department of Chemistry, Sapienza Universita di Roma, Piazzale Aldo Moro 5, 00185 
      Rome, Italy.
AD  - CNR NANOTEC, Campus Ecotekne, University of Salento, Via Monteroni, 73100 Lecce, 
      Italy.
FAU - Montone, Carmela Maria
AU  - Montone CM
AD  - Department of Chemistry, Sapienza Universita di Roma, Piazzale Aldo Moro 5, 00185 
      Rome, Italy.
FAU - Piovesana, Susy
AU  - Piovesana S
AUID- ORCID: 0000-0001-7134-7421
AD  - Department of Chemistry, Sapienza Universita di Roma, Piazzale Aldo Moro 5, 00185 
      Rome, Italy.
FAU - Zenezini Chiozzi, Riccardo
AU  - Zenezini Chiozzi R
AD  - Biomolecular Mass Spectrometry and Proteomics, Bijvoet Center for Biomolecular 
      Research and Utrecht Institute for Pharmaceutical Sciences, Utrecht University, 
      Padualaan 8, Utrecht 3584 CH, Netherlands.
AD  - Netherlands Proteomics CentrePadualaan 8Utrecht3584 CHNetherlands.
FAU - Cavaliere, Chiara
AU  - Cavaliere C
AUID- ORCID: 0000-0003-1332-682X
AD  - Department of Chemistry, Sapienza Universita di Roma, Piazzale Aldo Moro 5, 00185 
      Rome, Italy.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20200520
PL  - United States
TA  - Anal Chem
JT  - Analytical chemistry
JID - 0370536
RN  - 0 (Ferric Compounds)
RN  - 0 (Peptides)
RN  - 15FIX9V2JP (titanium dioxide)
RN  - D1JT611TNE (Titanium)
SB  - IM
MH  - Chromatography, High Pressure Liquid
MH  - Ferric Compounds/chemistry
MH  - Peptides/*analysis/metabolism
MH  - *Proteomics
MH  - Titanium/chemistry
EDAT- 2020/05/10 06:00
MHDA- 2021/02/13 06:00
CRDT- 2020/05/09 06:00
PHST- 2020/05/10 06:00 [pubmed]
PHST- 2021/02/13 06:00 [medline]
PHST- 2020/05/09 06:00 [entrez]
AID - 10.1021/acs.analchem.0c01342 [doi]
PST - ppublish
SO  - Anal Chem. 2020 Jun 2;92(11):7964-7971. doi: 10.1021/acs.analchem.0c01342. Epub 
      2020 May 20.