Screening for proteolytic activities in snake venom by means of a multiplexing electrospray ionization mass spectrometry assay scheme
| Autor: | Marianne Wilmer, Reto Schöni, André Liesener, Anna-Maria Perchuc, Uwe Karst |
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| Rok vydání: | 2005 |
| Předmět: |
Spectrometry
Mass Electrospray Ionization Electrospray ionization Enzyme-Linked Immunosorbent Assay Venom Mass spectrometry Peptide Mapping Substrate Specificity Analytical Chemistry Bothrops moojeni Crotalid Venoms Animals Humans Bothrops Multiplex Spectroscopy chemistry.chemical_classification Chromatography biology Organic Chemistry Substrate (chemistry) biology.organism_classification Enzyme Activation Enzyme chemistry Snake venom Cattle Peptide Hydrolases |
| Zdroj: | Rapid Communications in Mass Spectrometry. 19:2923-2928 |
| ISSN: | 1097-0231 0951-4198 |
| DOI: | 10.1002/rcm.2136 |
| Popis: | A multiplexed mass spectrometry based assay scheme for the simultaneous determination of five different substrate/product pairs was developed as a tool for screening of proteolytic activities in snake venom fractions from Bothrops moojeni. The assay scheme was employed in the functional characterization of eight model proteases. Time-resolved reaction profiles were generated and the relative reaction progress at each time point was determined. These were used to semi-quantitatively sort the catalytic activities of each enzyme towards the respective substrates into six classes. The resulting activity pattern served as an activity fingerprint for each enzyme. The multiplex assay scheme was then applied to a screening for proteolytic activities in fractions of the pre-separated venom from B. moojeni. Activity patterns of each fraction were generated and used to sort the fractions into three different categories of activity. By comparison of the fingerprint activity patterns of the venom fractions and the model enzymes, a compound with proteolytic properties similar to activated protein C was detected. |
| Databáze: | OpenAIRE |
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