A simple and efficient approach for preparing cationic coating with tunable electroosmotic flow for capillary zone electrophoresis-mass spectrometry-based top-down proteomics.

Electrophoresis, Capillary / methods Proteomics / methods Electroosmosis Cations / chemistry Tandem Mass Spectrometry / methods Saccharomyces cerevisiae / chemistry
Capillary zone electrophoresis-mass spectrometry Covalent cationic coating Large proteoforms Proteoform Reproducibility Top-down proteomics

Journal

Analytica chimica acta
ISSN: 1873-4324
Titre abrégé: Anal Chim Acta
Pays: Netherlands
ID NLM: 0370534

Informations de publication

Date de publication:
01 Nov 2024
Historique:
received: 12 06 2024
revised: 20 08 2024
accepted: 26 08 2024
medline: 13 9 2024
pubmed: 13 9 2024
entrez: 12 9 2024
Statut: ppublish

Résumé

Capillary zone electrophoresis-tandem mass spectrometry (CZE-MS/MS) has become a valuable analytical technique in top-down proteomics (TDP). CZE-MS/MS-based TDP typically employs separation capillaries with neutral coatings (i.e., linear polyacrylamide, LPA). However, issues related to separation resolution and reproducibility remain with the LPA-coated capillaries due to the unavoidable non-specific protein adsorption onto the capillary wall. Cationic coatings can be critical alternatives to LPA coating for CZE-MS/MS-based TDP due to the electrostatic repulsion between the positively charged capillary inner wall and proteoform molecules in the acidic separation buffer. Unfortunately, there are only very few studies using cationic coating-based CZE-MS/MS for TDP studies. In this work, we aimed to develop a simple and efficient approach for preparing separation capillaries with a cationic coating, i.e., poly (acrylamide-co-(3-acrylamidopropyl) trimethylammonium chloride [PAMAPTAC]) for CZE-MS/MS-based TDP. The PAMAPTAC coating-based CZE-MS produced significantly better separation resolution of proteoforms compared to the traditionally used LPA-coated approach. It achieved reproducible separation and measurement of a simple proteoform mixture and a complex proteome sample (i.e., a yeast cell lysate) regarding migration time, proteoform intensity, and the number of proteoform identifications. The PAMAPTAC coating-based CZE-MS enabled the detection of large proteoforms (≥30 kDa) from the yeast cell lysate reproducibly without any size-based prefractionation. Interestingly, the mobility of proteoforms using the PAMAPTAC coating can be predicted accurately using a simple semi-empirical model. The results render the PAMAPTAC coating as a valuable alternative to the LPA coating to advance CZE-MS-based TDP towards high-resolution separation and highly reproducible measurement of proteoforms in complex samples.

Sections du résumé

BACKGROUND BACKGROUND
Capillary zone electrophoresis-tandem mass spectrometry (CZE-MS/MS) has become a valuable analytical technique in top-down proteomics (TDP). CZE-MS/MS-based TDP typically employs separation capillaries with neutral coatings (i.e., linear polyacrylamide, LPA). However, issues related to separation resolution and reproducibility remain with the LPA-coated capillaries due to the unavoidable non-specific protein adsorption onto the capillary wall. Cationic coatings can be critical alternatives to LPA coating for CZE-MS/MS-based TDP due to the electrostatic repulsion between the positively charged capillary inner wall and proteoform molecules in the acidic separation buffer. Unfortunately, there are only very few studies using cationic coating-based CZE-MS/MS for TDP studies.
RESULTS RESULTS
In this work, we aimed to develop a simple and efficient approach for preparing separation capillaries with a cationic coating, i.e., poly (acrylamide-co-(3-acrylamidopropyl) trimethylammonium chloride [PAMAPTAC]) for CZE-MS/MS-based TDP. The PAMAPTAC coating-based CZE-MS produced significantly better separation resolution of proteoforms compared to the traditionally used LPA-coated approach. It achieved reproducible separation and measurement of a simple proteoform mixture and a complex proteome sample (i.e., a yeast cell lysate) regarding migration time, proteoform intensity, and the number of proteoform identifications. The PAMAPTAC coating-based CZE-MS enabled the detection of large proteoforms (≥30 kDa) from the yeast cell lysate reproducibly without any size-based prefractionation. Interestingly, the mobility of proteoforms using the PAMAPTAC coating can be predicted accurately using a simple semi-empirical model.
SIGNIFICANCE CONCLUSIONS
The results render the PAMAPTAC coating as a valuable alternative to the LPA coating to advance CZE-MS-based TDP towards high-resolution separation and highly reproducible measurement of proteoforms in complex samples.

Identifiants

DOI: 10.1016/j.aca.2024.343162 PMID: 39266194
pubmed: 39266194
pii: S0003-2670(24)00963-2
doi: 10.1016/j.aca.2024.343162
pii:
doi:

Substances chimiques

Cations 0

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

343162

Informations de copyright

Copyright © 2024 Elsevier B.V. All rights reserved.

Déclaration de conflit d'intérêts

Declaration of competing interest The authors declare no competing financial interest.

Auteurs

Qianjie Wang (Q)

Department of Chemistry, Michigan State University, 578 S Shaw Lane, East Lansing, MI, 48824, USA; Department of Biochemistry and Molecular Biology, Michigan State University, East Lansing, MI, 48824, USA; Plant Resilience Institute, Michigan State University, East Lansing, MI, 48824, USA.

Guangyao Gao (G)

Department of Chemistry, Michigan State University, 578 S Shaw Lane, East Lansing, MI, 48824, USA.

Fei Fang (F)

Department of Chemistry, Michigan State University, 578 S Shaw Lane, East Lansing, MI, 48824, USA.

Qianyi Wang (Q)

Department of Chemistry, Michigan State University, 578 S Shaw Lane, East Lansing, MI, 48824, USA.

Peter K Lundquist (PK)

Department of Biochemistry and Molecular Biology, Michigan State University, East Lansing, MI, 48824, USA; Plant Resilience Institute, Michigan State University, East Lansing, MI, 48824, USA.

Liangliang Sun (L)

Department of Chemistry, Michigan State University, 578 S Shaw Lane, East Lansing, MI, 48824, USA. Electronic address: lsun@chemistry.msu.edu.

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Classifications MeSH

questionsmedicales.fr › Techniques d'investigation › Techniques électrochimiques › Électrophorèse › Électrophorèse capillaire › A simple and efficient approach for preparing...
questionsmedicales.fr › Disciplines des sciences naturelles › Chimie › Biochimie › Protéomique › A simple and efficient approach for preparing...
questionsmedicales.fr › Phénomènes chimiques › Perméabilité › Osmose › Électro-osmose › A simple and efficient approach for preparing...
questionsmedicales.fr › Produits chimiques inorganiques › Électrolytes › Ions › Cations › A simple and efficient approach for preparing...
questionsmedicales.fr › Techniques d'investigation › Techniques de chimie analytique › Spectrométrie de masse › Spectrométrie de masse en tandem › A simple and efficient approach for preparing...
questionsmedicales.fr › Eucaryotes › Champignons › Levures › Saccharomyces › Saccharomyces cerevisiae › A simple and efficient approach for preparing...