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. 2022 Dec 1;27(23):8406.
doi: 10.3390/molecules27238406.

Quantitative Assessment of Serine-8 Phosphorylated β-Amyloid Using MALDI-TOF Mass Spectrometry

Andrey A Kuzin  1 , Galina S Stupnikova  1 , Polina A Strelnikova  2   3 , Ksenia V Danichkina  1 , Maria I Indeykina  2   4 , Stanislav I Pekov  1   3   4   5 , Igor A Popov  1   5
Affiliations

Quantitative Assessment of Serine-8 Phosphorylated β-Amyloid Using MALDI-TOF Mass Spectrometry

Andrey A Kuzin et al. Molecules. .

Abstract

The study of the molecular mechanisms of the pathogenesis of Alzheimer's disease (AD) is extremely important for identifying potential therapeutic targets as well as early markers. In this regard, the study of the role of post-translational modifications (PTMs) of β-amyloid (Aβ) peptides is of particular relevance. Serine-8 phosphorylated forms (pSer8-Aβ) have been shown to have an increased aggregation capacity and may reflect the severity of amyloidosis. Here, an approach for quantitative assessment of pSer8-Aβ based on matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is proposed. The relative fraction of pSer8-Aβ was estimated in the total Aβ-pool with a detection limit of 1 fmol for pSer8-Aβ (1-16) and an accuracy of 2% for measurements in the reflectron mode. The sensitivity of the developed method is suitable for determining the proportion of phosphorylated peptides in biological samples.

Keywords: MALDI-TOF; amyloid-beta; phosphopeptide.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Comparison of mass spectra of pSer8-Aβ(1–16) in linear and reflectron modes. The blue line shows the mass spectrum of pSer8-Aβ(1–16) in reflectron mode, orange—Aβ(1–16) in reflectron mode, and red—pSer8-Aβ(1–16) in linear mode.
Figure 2
Figure 2
Ion map for pSer8-Aβ16 mass spectra obtained using DHAP, HCCA, and DHB matrices with the addition of various concentrations of phosphoric and trifluoroacetic acid. Aβ16 corresponds to m/z 1955; pSer8-Aβ16 corresponds to m/z 2035.
Figure 3
Figure 3
Calibration curve for Aβ16 in linear mode on an AnchorChip target with 10 mg/mL DHB, 0.1% TFA.
Figure 4
Figure 4
Calibration curve for Aβ16 in reflectron mode on an AnchorChip target with 10 mg/mL DHB, 0.1% TFA.
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