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Positive Effect of Acetylation on Proteomic Analysis Based on Liquid Chromatography with Atmospheric Pressure Chemical Ionization and Photoionization Mass Spectrometry
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SYSNO ASEP 0572296 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Positive Effect of Acetylation on Proteomic Analysis Based on Liquid Chromatography with Atmospheric Pressure Chemical Ionization and Photoionization Mass Spectrometry Author(s) Sedláčková, Simona (UOCHB-X)
Hubálek, Martin (UOCHB-X) RID, ORCID
Vrkoslav, Vladimír (UOCHB-X) RID, ORCID
Blechová, Miroslava (UOCHB-X)
Kozlík, P. (CZ)
Cvačka, Josef (UOCHB-X) RID, ORCIDArticle number 3711 Source Title Molecules. - : MDPI
Roč. 28, č. 9 (2023)Number of pages 20 s. Language eng - English Country CH - Switzerland Keywords chemical ionization ; photoionization ; peptides acetylation OECD category Analytical chemistry R&D Projects GA20-09126S GA ČR - Czech Science Foundation (CSF) Method of publishing Open access Institutional support UOCHB-X - RVO:61388963 UT WOS 000987656600001 EID SCOPUS 85159358512 DOI 10.3390/molecules28093711 Annotation A typical bottom-up proteomic workflow comprises sample digestion with trypsin, separation of the hydrolysate using reversed-phase HPLC, and detection of peptides via electrospray ionization (ESI) tandem mass spectrometry. Despite the advantages and wide usage of protein identification and quantification, the procedure has limitations. Some domains or parts of the proteins may remain inadequately described due to inefficient detection of certain peptides. This study presents an alternative approach based on sample acetylation and mass spectrometry with atmospheric pressure chemical ionization (APCI) and atmospheric pressure photoionization (APPI). These ionizations allowed for improved detection of acetylated peptides obtained via chymotrypsin or glutamyl peptidase I (Glu-C) digestion. APCI and APPI spectra of acetylated peptides often provided sequence information already at the full scan level, while fragmentation spectra of protonated molecules and sodium adducts were easy to interpret. As demonstrated for bovine serum albumin, acetylation improved proteomic analysis. Compared to ESI, gas-phase ionizations APCI and APPI made it possible to detect more peptides and provide better sequence coverages in most cases. Importantly, APCI and APPI detected many peptides which passed unnoticed in the ESI source. Therefore, analytical methods based on chymotrypsin or Glu-C digestion, acetylation, and APPI or APCI provide data complementary to classical bottom-up proteomics. Workplace Institute of Organic Chemistry and Biochemistry Contact asep@uochb.cas.cz ; Kateřina Šperková, Tel.: 232 002 584 ; Jana Procházková, Tel.: 220 183 418 Year of Publishing 2024 Electronic address https://doi.org/10.3390/molecules28093711
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