Projects per year
Abstract
Leucyl aminopeptidases (LAPs) are involved in multiple cellular functions, which, in the case of infectious diseases, includes participation in the pathogen-host cell interface and pathogenesis. Thus, LAPs are considered good candidate drug targets, and the major M17-LAP from Trypanosoma cruzi (LAPTc) in particular is a promising target for Chagas disease. To exploit LAPTc as a potential target, it is essential to develop potent and selective inhibitors. To achieve this, we report a high-throughput screening method for LAPTc. Two methods were developed and optimized: a Leu-7-amido-4-methylcoumarin-based fluorogenic assay and a RapidFire mass spectrometry (RapidFire MS)-based assay using the LSTVIVR peptide as substrate. Compared with a fluorescence assay, the major advantages of the RapidFire MS assay are a greater signal-to-noise ratio as well as decreased consumption of enzyme. RapidFire MS was validated with the broad-spectrum LAP inhibitors bestatin (IC50 = 0.35 μM) and arphamenine A (IC50 = 15.75 μM). We suggest that RapidFire MS is highly suitable for screening for specific LAPTc inhibitors.
Original language | English |
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Pages (from-to) | 1064-1071 |
Number of pages | 8 |
Journal | SLAS Discovery |
Volume | 25 |
Issue number | 9 |
Early online date | 13 May 2020 |
DOIs | |
Publication status | Published - 1 Oct 2020 |
Keywords
- arphamenine
- bestatin
- high-throughput screening
- M17-leucyl aminopeptidase
- RapidFire mass spectrometry
ASJC Scopus subject areas
- Analytical Chemistry
- Molecular Medicine
- Biochemistry
- Biotechnology
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Dive into the research topics of 'Development of a High-Throughput Screening Assay to Identify Inhibitors of the Major M17-Leucyl Aminopeptidase from Trypanosoma cruzi Using RapidFire Mass Spectrometry'. Together they form a unique fingerprint.Projects
- 2 Active
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A Systems Approach for Understanding Cell Surface Dynamics in Trypanosomes (Investigator Award)
1/10/17 → 31/03/24
Project: Research
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Wellcome Centre for Anti-Infectives Research
Cook, S., De Rycker, M., Fairlamb, A., Ferguson, M., Field, M., Gilbert, I., Gray, D., Horn, D., Pawlowic, M., Read, K., Wyatt, P. & Wyllie, S.
1/04/17 → 31/03/24
Project: Research