Ion soft-landing into liquids: Protein identification, separation, and purification with retention of biological activity
| Autor: | Jormarie Alvarez, Justin M. Wiseman, R. Graham Cooks, Nari Talaty, Zheng Ouyang, Bogdan Gologan, Zoltan Takats |
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| Rok vydání: | 2004 |
| Předmět: |
Ions
Spectrometry Mass Electrospray Ionization Chromatography Molecular Sequence Data Proteins Fast atom bombardment Mass spectrometry chemistry.chemical_compound Matrix-assisted laser desorption/ionization chemistry Structural Biology Hexokinase Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization Desorption Monolayer Protein purification Glycerol Muramidase Amino Acid Sequence Lysozyme Peptides Spectroscopy |
| Zdroj: | Journal of the American Society for Mass Spectrometry. 15:1874-1884 |
| ISSN: | 1044-0305 |
| DOI: | 10.1016/j.jasms.2004.09.005 |
| Popis: | Protein ions, after mass spectrometric separation, can be soft-landed into liquid surfaces with preservation of their native structures. Retention of biological activity is strongly favored in glycerol-based surfaces but not in self-assembled monolayer solid surfaces. Soft-landing efficiency for multiply-charged hexokinase ions was found to be some four times higher for a glycerol/fructose liquid surface than for a fluorinated self-assembled monolayer surface. Soft-landing into liquid surfaces is also shown to allow (1) protein purification, (2) on-surface identification of the soft-landed material using MALDI, and (3) protein identification by in-surface tryptic digestion. Pure lysozyme was successfully isolated from different mixtures including an oxidized, partially decomposed batch of the protein and a partial tryptic digest. Liquid glycerol/carbohydrate mixtures could be used directly to record MALDI spectra on the soft-landed compounds provided they were fortified in advance with traditional MALDI matrices such as p-nitroaniline and alpha-cyano-4-hydroxycinnamic acid. Various proteins were soft-landed and detected on-target using these types of liquid surface. Soft-landing of multiply-charged lysozyme ions onto fluorinated self-assembled monolayer surfaces was found to occur with a limited amount of neutralization, and trapped multiply-charged ions could be desorbed from the surface by laser desorption. Initial data is shown for a new approach to protein identification that combines top-down and bottom-up approaches by utilizing protein ion soft-landing from a protein mixture, followed by tryptic digestion of the landed material and detection of characteristic tryptic fragments by MALDI. |
| Databáze: | OpenAIRE |
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