Protein identification based on matrix assisted laser desorption/ionization-post source decay-mass spectrometry
| Autor: | Kris Gevaert, Marc Goethals, Bart Hoorelbeke, Hans Demol, Joël Vandekerckhove, Magda Puype, Lennart Martens, Jozef Van Damme, Stefaan De Boeck |
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| Rok vydání: | 2001 |
| Předmět: |
MALDI imaging
Chromatography Protein mass spectrometry Chemistry Clinical Biochemistry Analytical chemistry Proteins Mass spectrometry Biochemistry Capillary electrophoresis–mass spectrometry Sample preparation in mass spectrometry Peptide Fragments Analytical Chemistry Surface-enhanced laser desorption/ionization Matrix-assisted laser desorption/ionization Peptide mass fingerprinting Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization |
| Zdroj: | Electrophoresis. 22(9) |
| ISSN: | 0173-0835 |
| Popis: | Due to its very short analysis time, its high sensitivity and ease of automation, matrix-assisted laser desorption/ionization (MALDI)-peptide mass fingerprinting has become the preferred method for identifying proteins of which the sequences are available in databases. However, many protein samples cannot be unambiguously identified by exclusively using their peptide mass fingerprints (e.g., protein mixtures, heavily posttranslationally modified proteins and small proteins). In these cases, additional sequence information is needed and one of the obvious choices when working with MALDI-mass spectrometry (MS) is to choose for post source decay (PSD) analysis on selected peptides. This can be performed on the same sample which is used for peptide mass fingerprinting. Although in this type of peptide analysis, fragmentation yields are very low and PSD spectra are often very difficult to interpret manually, we here report upon our five years of experience with the use of PSD spectra for protein identification in sequence (protein or expressed sequence tag (EST)) databases. The combination of peptide mass fingerprinting and PSD and analysis described here generally leads to unambiguous protein identification in the amount of material range generally encountered in most proteome studies. |
| Databáze: | OpenAIRE |
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