Improved Protein Coverage in Bottom-Up Proteomes Analysis Using Fluoroalcohol-Mediated Supramolecular Biphasic Systems With Mixed Amphiphiles for Sample Extraction, Fractionation, and Enrichment
| Autor: | Sajad Tasharofi, Durga D. Khanal, Morteza G. Khaledi, Mohammadmehdi Azizi |
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| Rok vydání: | 2021 |
| Předmět: |
Proteomics
Coacervate Chromatography Proteome Chemistry 010401 analytical chemistry Membrane Proteins Saccharomyces cerevisiae Fractionation Chemical Fractionation 010402 general chemistry 01 natural sciences 0104 chemical sciences Analytical Chemistry Membrane Membrane protein Pulmonary surfactant Integral membrane protein |
| Zdroj: | Analytical Chemistry. 93:7430-7438 |
| ISSN: | 1520-6882 0003-2700 |
| Popis: | A new class of supramolecular biphasic systems containing fluoroalcohol-induced coacervates (FAiC) provides concomitant fractionation of complex protein mixtures, high solubilizing power for extraction of various types of proteins, especially those with high hydrophobicity (such as membrane proteins), and enrichment of low-abundance proteins. Subsequently, the use of FAiC biphasic systems (BPS) in the bottom-up proteomics workflow resulted in significantly higher coverage for the whole proteome, various subproteomes, especially those embedded or associated with membranes, post-translationally modified proteins, and low-abundance proteins (LAPs) as compared to the conventional methodologies. In this work, we used a new type of FAiC-BPS composed of mixed amphiphiles, a zwitterionic surfactant 3-(N,N-dimethylmyristyl ammonia) propane sulfonate (DMMAPS), a quaternary ammonium salt (QUATS), and hexafluoroisopropanol (HFIP) as the coacervator for extraction, fractionation, and enrichment of yeast proteome in bottom-up proteomics. The coverage of the lower-abundance proteins (abundance below 2000 molecules/cell) improved by more than 100% using DMMAPS and DMMAPS + QUATS systems as compared to the conventional methods using urea or detergent solutions for protein solubilization. Additionally, these coacervate systems show increased coverage of integral membrane proteins and proteins with α-helices by up to 24 and 555%, respectively. |
| Databáze: | OpenAIRE |
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