Screening Assay for Metal-Catalyzed Oxidation Inhibitors Using Liquid Chromatography−Mass Spectrometry with an N-Terminal β-Amyloid Peptide
Abstract
Production of microregional catalytic reactive oxygen species (ROS) by metal-binding amyloid-beta (Abeta) peptides mediates the neurotoxicity of Alzheimer's disease, and inhibitors of this activity may be of therapeutic value. No current analytical methods target specific ROS inhibitors produced by metal-binding peptides. We report a screening assay for metal-catalyzed oxidation (MCO) inhibitors based on liquid chromatography-mass spectrometry (LC-MS) with a model N-terminal Abeta peptide (Abeta(1-6)). When subjected to MCO by Cu(II)/ascorbic acid, singly and doubly charged Abeta(1-6) molecules were observed at m/z 729.2 and 364.8 and m/z 685.3 and 343.3, respectively, corresponding to a decrease in mass of 45 and 89 Da compared with the model peptide. In contrast, H(2)O(2) did not modify the Abeta(1-6) peptide. Modified peptides were characterized by a specific MCO of Abeta(1-6), which contains both His and N-terminal Asp residues. LC-MS detection of the modified peptides allowed us to identify antioxidants that inhibit MCO of Abeta(1-6). MCO of the model peptide was inhibited by curcumin, but not dibutylhydroxytoluene, carotene, tocopherol, estradiol or nicotine, revealing a clear difference between curcumin and other antioxidants. This novel assay may allow for the identification of antioxidants that protect against MCO of peptides and proteins related to degenerative diseases.
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Both in vitro and in vivo studies pointed out the Aβ peptide as a target molecule of metal-catalyzed oxidation.
“Moreover, both in vitro and in vivo studies pointed out the Aβ peptide as a target molecule of metal-catalyzed oxidation (Näslund et al., 1994; Kowalik-Jankowska et al., 2004; Inoue et al., 2009).”
From Exploring the Therapeutic Potential of Phytochemicals in Alzheimer’s Disease: Focus on Polyphenols and Monoterpenes (2022) · cites this paper as Inoue et al., 2009
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- Authors
- Inoue, K., Nakagawa, A., Hino, T., & Oka, H.
- Affiliations
- Department of Physical and Analytical Chemistry, School of Pharmacy, and Graduate School of Human Ecology, Human Ecology Major, Kinjo Gakuin University, 2-1723 Omori, Moriyama-ku, Nagoya 463-8521, Japan
- Kinjo Gakuin University (JP)
- ORCID
- Oka, Hisao
- Venue
- Analytical Chemistry
- Publisher
- American Chemical Society (ACS)
- Published
- 2009-01-27
- Volume
- 81
- Issue
- 5
- Pages
- 1819-1825
- ISSN
- 0003-2700 (ISSN-L)
- 1520-6882
- OpenAlex
- W2063078119
- Cited by
- 32
- References
- 49
- Citations by year
- 2025: 1 · 2024: 2 · 2023: 2 · 2022: 2 · 2020: 2 · 2019: 3 · 2017: 5 · 2016: 3 · 2013: 3 · 2012: 2
- FWCI
- 1.99
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Analytical Chemistry (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Health Sciences › Medicine › Physiology › Alzheimer's disease research and treatments
- Physical Sciences › Chemistry › Spectroscopy › Mass Spectrometry Techniques and Applications
- Physical Sciences › Chemistry › Organic Chemistry › Free Radicals and Antioxidants
- Keywords
- metal-catalyzed oxidation, amyloid-beta, reactive oxygen species, Alzheimer's disease, antioxidants, curcumin, copper(II) catalysis, vitamin C, peptide oxidation, neurotoxicity, screening assay
- MeSH terms
- Catalysis, Chromatography, Liquid, Drug Evaluation, Preclinical, Hydrogen Peroxide, Metals, Oxidation-Reduction, Peptide Fragments, Mass Spectrometry, Amyloid beta-Peptides, Reactive Oxygen Species
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