Fructose and methylglyoxal-induced glycation alters structural and functional properties of salivary proteins, albumin and lysozyme

Muraoka, Mariane Yumiko; Justino, Allisson Benatti; Caixeta, Douglas Carvalho; Queiroz, Julia Silveira; Sabino-Silva, Robinson; Espíndola, Foued Salmen

doi:10.1371/journal.pone.0262369

Cited by 18 publications

(7 citation statements)

References 122 publications

(133 reference statements)

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“…Spectra were acquired with 4 cm −1 spectral resolution and 32 scans per sample, from 400 to 4000 cm −1 . Two replicates per sample were studied and the mean was used for each sample [ 21 , 23 , 31 ].…”

Section: Methodsmentioning

confidence: 99%

Salivary Detection of Zika Virus Infection Using ATR-FTIR Spectroscopy Coupled with Machine Learning Algorithms and Univariate Analysis: A Proof-of-Concept Animal Study

et al. 2023

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Zika virus (ZIKV) diagnosis is currently performed through an invasive, painful, and costly procedure using molecular biology. Consequently, the search for a non-invasive, more cost-effective, reagent-free, and sustainable method for ZIKV diagnosis is of great relevance. It is critical to prepare a global strategy for the next ZIKV outbreak given its devastating consequences, particularly in pregnant women. Attenuated total reflection–Fourier transform infrared (ATR-FTIR) spectroscopy has been used to discriminate systemic diseases using saliva; however, the salivary diagnostic application in viral diseases is unknown. To test this hypothesis, we intradermally challenged interferon-gamma gene knockout C57/BL6 mice with ZIKV (50 µL,105 FFU, n = 7) or vehicle (50 µL, n = 8). Saliva samples were collected on day three (due to the peak of viremia) and the spleen was also harvested. Changes in the salivary spectral profile were analyzed by Student’s t test (p < 0.05), multivariate analysis, and the diagnostic capacity by ROC curve. ZIKV infection was confirmed by real-time PCR of the spleen sample. The infrared spectroscopy coupled with univariate analysis suggested the vibrational mode at 1547 cm−1 as a potential candidate to discriminate ZIKV and control salivary samples. Three PCs explained 93.2% of the cumulative variance in PCA analysis and the spectrochemical analysis with LDA achieved an accuracy of 93.3%, with a specificity of 87.5% and sensitivity of 100%. The PCA-SVM analysis showed 100% discrimination between both classes. Our results suggest that ATR-FTIR applied to saliva might have high accuracy in ZIKV diagnosis with potential as a non-invasive and cost-effective diagnostic tool.

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Section: Methodsmentioning

confidence: 99%

Salivary Detection of Zika Virus Infection Using ATR-FTIR Spectroscopy Coupled with Machine Learning Algorithms and Univariate Analysis: A Proof-of-Concept Animal Study

et al. 2023

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“…In this study, we focused on the formation of fluorescent AGEs by several proteins other than BSA that have been reported to undergo glycation [37][38][39][40][41]. Using MGO as a glycation agent expected to have higher reactivity than glucose, we attempted to establish a method for evaluating the activity of antiglycation compounds based on the formation of ARP, a representative fluorescent AGE, and the inhibition of its formation.…”

Section: Discussionmentioning

confidence: 99%

Novel Fluorometric Assay of Antiglycation Activity Based on Methylglyoxal-Induced Protein Carbonylation

Koike,

Saito,

Ogasawara

2023

Antioxidants

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Advanced glycation end products (AGEs), which can have multiple structures, are formed at the sites where the carbonyl groups of reducing sugars bind to the free amino groups of proteins through the Maillard reaction. Some AGE structures exhibit fluorescence, and this fluorescence has been used to measure the formation and quantitative changes in carbonylated proteins. Recently, fluorescent AGEs have also been used as an index for the evaluation of compounds that inhibit protein glycation. However, the systems used to generate fluorescent AGEs from the reaction of reducing sugars and proteins used for the evaluation of antiglycation activity have not been determined through appropriate research; thus, problems remain regarding sensitivity, quantification, and precision. In the present study, using methylglyoxal (MGO), a reactive carbonyl compound to induce glycation, a comparative analysis of the mechanisms of formation of fluorescent substances from several types of proteins was conducted. The analysis identified hen egg lysozyme (HEL) as a protein that produces stronger fluorescent AGEs faster in the Maillard reaction with MGO. It was also found that the AGE structure produced in MGO-induced in HEL was argpyrimidine. By optimizing the reaction system, we developed a new evaluation method for compounds with antiglycation activity and established an efficient evaluation method (HEL–MGO assay) with greater sensitivity and accuracy than the conventional method, which requires high concentrations of bovine serum albumin and glucose. Furthermore, when compounds known to inhibit glycation were evaluated using this method, their antiglycation activities were clearly and significantly measured, demonstrating the practicality of this method.

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“…Fluorescent spectroscopy is usually used to monitor the rate of formation of AGEs, considering that most of the advanced products of glycation are fluorophores [ 140 ]. The rate of these reactions is quite slow, and only proteins with large amounts of lysine residues undergo glycation, with significant amounts of AGEs leading to reduced protein functionality and stability [ 141 , 142 ].…”

Section: Glycosylationmentioning

confidence: 99%

“…It is known that proteins from saliva show a very quick turnover; nevertheless, few studies point out how the loss of functional activity of some salivary proteins is to be related to glycation processes. The antibacterial activity of lactoferrin or lysozyme and the antioxidant capacity of superoxide dismutase, all detectable in human saliva, decreased after the formation of AGEs [ 142 , 145 , 146 ].…”

Section: Glycosylationmentioning

confidence: 99%

The Post-Translational Modifications of Human Salivary Peptides and Proteins Evidenced by Top-Down Platforms

Messana

Manconi

Cabras

et al. 2023

IJMS

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In this review, we extensively describe the main post-translational modifications that give rise to the multiple proteoforms characterized to date in the human salivary proteome and their potential role. Most of the data reported were obtained by our group in over twenty-five years of research carried out on human saliva mainly by applying a top-down strategy. In the beginning, we describe the products generated by proteolytic cleavages, which can occur before and after secretion. In this section, the most relevant families of salivary proteins are also described. Next, we report the current information concerning the human salivary phospho-proteome and the limited news available on sulfo-proteomes. Three sections are dedicated to the description of glycation and enzymatic glycosylation. Citrullination and N- and C-terminal post-translational modifications (PTMs) and miscellaneous other modifications are described in the last two sections. Results highlighting the variation in the level of some proteoforms in local or systemic pathologies are also reviewed throughout the sections of the manuscript to underline the impact and relevance of this information for the development of new diagnostic biomarkers useful in clinical practice.

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Fructose and methylglyoxal-induced glycation alters structural and functional properties of salivary proteins, albumin and lysozyme

Cited by 18 publications

References 122 publications

Salivary Detection of Zika Virus Infection Using ATR-FTIR Spectroscopy Coupled with Machine Learning Algorithms and Univariate Analysis: A Proof-of-Concept Animal Study

Salivary Detection of Zika Virus Infection Using ATR-FTIR Spectroscopy Coupled with Machine Learning Algorithms and Univariate Analysis: A Proof-of-Concept Animal Study

Novel Fluorometric Assay of Antiglycation Activity Based on Methylglyoxal-Induced Protein Carbonylation

The Post-Translational Modifications of Human Salivary Peptides and Proteins Evidenced by Top-Down Platforms

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