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EThcD and 213 nm UVPD for Top-Down Analysis of Bovine Seminal Plasma Proteoforms on Electrophoretic and Chromatographic Time Frames
- Source :
- Analytical Chemistry. 92:2979-2987
- Publication Year :
- 2020
- Publisher :
- American Chemical Society (ACS), 2020.
-
Abstract
- Seminal plasma is a critical and complex fluid that carries sperm to eggs to initiate the fertilization process. Here, we present a top-down mass spectrometry (TDMS) strategy for identifying proteins and posttranslational modifications (PTMs) in bovine seminal plasma. In this study, proteins were separated using sheathless capillary zone electrophoresis (CZE)-MS and reversed-phase liquid chromatography (LC)-MS, and then fragmented using electron-transfer/higher-energy collisional dissociation (EThcD) and 213 nm ultraviolet photodissociation (213 nm UVPD) to provide more comprehensive information about the proteomic landscape of this biological fluid. Four hundred and seventeen proteoforms were identified by sheathless CZE-MS, and one hundred and seventy-two species were unique to this method. LC-MS identified 3090 proteoforms, including 1707 unique species. All identifications were within ±10 ppm (mass error) and with a P-Score ≤1 × 10–04. Pooling results (triplicate measurements) from sheathless CZE-MS a...
- Subjects :
- Chromatography, Reverse-Phase
Chromatography
Ultraviolet Rays
Chemistry
010401 analytical chemistry
Plasma
010402 general chemistry
Mass spectrometry
01 natural sciences
Mass Spectrometry
Biological fluid
0104 chemical sciences
Analytical Chemistry
Electron Transport
Electrophoresis
Capillary electrophoresis
Semen
Top down analysis
Animals
Cattle
Subjects
Details
- ISSN :
- 15206882 and 00032700
- Volume :
- 92
- Database :
- OpenAIRE
- Journal :
- Analytical Chemistry
- Accession number :
- edsair.doi.dedup.....f888771b271e39cd0384526b21a6e6ba