Native GELFrEE: A New Separation Technique for Biomolecular Assemblies
| Autor: | Owen S. Skinner, Marcelo Valle de Sousa, Neil L. Kelleher, Luis H. F. Do Vale, Pierre C. Havugimana, Philip D. Compton, Adam D. Catherman |
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| Rok vydání: | 2015 |
| Předmět: |
Proteomics
Swine Molecular Sequence Data Stacking High resolution Fractionation Chemical Fractionation Mass spectrometry Mass Spectrometry Analytical Chemistry Rats Sprague-Dawley Mice Animals Humans Amino Acid Sequence Trichoderma Chromatography Chemistry Chemical fractionation Proteins Heart Rats Sprague dawley Multiprotein Complexes Electrophoresis Polyacrylamide Gel Biological system Multistage mass spectrometry |
| Zdroj: | Analytical Chemistry. 87:3032-3038 |
| ISSN: | 1520-6882 0003-2700 |
| Popis: | The cadre of protein complexes in cells performs an array of functions necessary for life. Their varied structures are foundational to their ability to perform biological functions, lending great import to the elucidation of complex composition and dynamics. Native separation techniques that are operative on low sample amounts and provide high resolution are necessary to gain valuable data on endogenous complexes. Here, we detail and optimize the use of tube gel separations to produce samples proven compatible with native, multistage mass spectrometry (nMS/MS). We find that a continuous system (i.e., no stacking gel) with a gradient in its extent of cross-linking and use of the clear native buffer system performs well for both fractionation and native mass spectrometry of heart extracts and a fungal secretome. This integrated advance in separations and nMS/MS offers the prospect of untargeted proteomics at the next hierarchical level of protein organization in biology. |
| Databáze: | OpenAIRE |
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