Untargeted adductomics of Cys34 modifications to human serum albumin in newborn dried blood spots

Yano, Yukiko; Grigoryan, Hasmik; Schiffman, Courtney; Edmands, William M. B.; Petrick, Lauren; Hall, Katie; Whitehead, Todd P.; Metayer, Catherine; Dudoit, Sandrine; Rappaport, Stephen M.

doi:10.1007/s00216-019-01675-8

Cited by 25 publications

(23 citation statements)

References 63 publications

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“…The peak area ratio (PAR = adduct peak area/HK peptide peak area) has been shown to be a reliable measure of adduct concentration over a wide dynamic range [36]. Recently the method has been adapted for analysis of HSA extracted from dried blood spots (DBS), which require additional steps prior to digestion [38]. The small amount of protein needed for HSA adductomics, as well as the possibility to use DBS for the analysis, is an attractive advantage of this methodology.…”

Section: Sample Preparationmentioning

confidence: 99%

Protein Adductomics: Methodologies for Untargeted Screening of Adducts to Serum Albumin and Hemoglobin in Human Blood Samples

2019

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The reaction products of electrophiles in vivo can be measured as adducts to the abundant proteins, hemoglobin (Hb), and human serum albumin (HSA), in human blood samples. During the last decade, methods for untargeted screening of such adducts, called “adductomics”, have used liquid chromatography-mass spectrometry to detect large numbers of previously unknown Hb and HSA adducts. This review presents methodologies that were developed and used in our laboratories for Hb and HSA adductomics, respectively. We discuss critical aspects regarding choice of target protein, sample preparation, mass spectrometry, data evaluation, and strategies for identification of detected unknown adducts. With this review we give an overview of these two methodologies used for protein adductomics and the precursor electrophiles that have been elucidated from the adducts.

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Section: Sample Preparationmentioning

confidence: 99%

Protein Adductomics: Methodologies for Untargeted Screening of Adducts to Serum Albumin and Hemoglobin in Human Blood Samples

2019

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“…A total of 656 NBS punches were analyzed (Supplementary Table 1). Briefly, samples were extracted with water and assayed for potassium [8] (batch 1) or hemoglobin [14] (batches 2-4) to adjust for blood volume (see Supplementary Figure 1). Then, acetonitrile was added to precipitate proteins and extracts were analyzed by LC-HRMS [8].…”

Section: Metabolomic Analysismentioning

confidence: 99%

Metabolomics of neonatal blood spots reveal distinct phenotypes of pediatric acute lymphoblastic leukemia and potential effects of early-life nutrition

et al. 2019

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Early-life exposures are believed to influence the incidence of pediatric acute lymphoblastic leukemia (ALL). Archived neonatal blood spots (NBS), collected within the first days of life, offer a means to investigate small molecules that reflect early-life exposures. Using untargeted metabolomics, we compared abundances of small-molecule features in extracts of NBS punches from 332 children that later developed ALL and 324 healthy controls. Subjects were stratified by early (1-5 y) and late (6-14 y) diagnosis. Mutually-exclusive sets of metabolic featuresrepresenting putative lipids and fatty acids-were associated with ALL, including 9 and 19 metabolites in the early-and late-diagnosis groups, respectively. In the late-diagnosis group, a prominent cluster of features with apparent 18:2 fatty-acid chains suggested that newborn exposure to the essential nutrient, linoleic acid, increased ALL risk. Interestingly, abundances of these putative 18:2 lipids were greater in infants who were fed formula rather than breast milk (colostrum) and increased with the mother's pre-pregnancy body mass index. These results suggest possible etiologic roles of newborn nutrition in late-diagnosis ALL.

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“…The use of thiol-affinity resins [30] and antibodies [31] was initially proposed for the enrichment of Cys34 adducts. However, more recent procedures do not use these pre-enrichment steps [32,33], the HSA is precipitated from serum with 60% methanol, and following trypsin digestion and prior to MS analysis an isotopically labeled peptide is added, as an internal standard, enabling corrections of retention time drifts and mass accuracy. Uchida´s group proposed a methodology based on the acid hydrolysis of proteins, such as low-density lipoproteins [34] and Hb [35], to amino acids for the comprehensive analysis of His and Lys adducts formed with lipid peroxidation products.…”

Section: Sample Pre-treatment and Adducts Enrichmentmentioning

confidence: 99%

Mass Spectrometry-Based Methodologies for Targeted and Untargeted Identification of Protein Covalent Adducts (Adductomics): Current Status and Challenges

et al. 2019

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Protein covalent adducts formed upon exposure to reactive (mainly electrophilic) chemicals may lead to the development of a wide range of deleterious health outcomes. Therefore, the identification of protein covalent adducts constitutes a huge opportunity for a better understanding of events underlying diseases and for the development of biomarkers which may constitute effective tools for disease diagnosis/prognosis, for the application of personalized medicine approaches and for accurately assessing human exposure to chemical toxicants. The currently available mass spectrometry (MS)-based methodologies, are clearly the most suitable for the analysis of protein covalent modifications, providing accuracy, sensitivity, unbiased identification of the modified residue and conjugates along with quantitative information. However, despite the huge technological advances in MS instrumentation and bioinformatics tools, the identification of low abundant protein covalent adducts is still challenging. This review is aimed at summarizing the MS-based methodologies currently used for the identification of protein covalent adducts and the strategies developed to overcome the analytical challenges, involving not only sample pre-treatment procedures but also distinct MS and data analysis approaches.

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Untargeted adductomics of Cys34 modifications to human serum albumin in newborn dried blood spots

Cited by 25 publications

References 63 publications

Protein Adductomics: Methodologies for Untargeted Screening of Adducts to Serum Albumin and Hemoglobin in Human Blood Samples

Protein Adductomics: Methodologies for Untargeted Screening of Adducts to Serum Albumin and Hemoglobin in Human Blood Samples

Metabolomics of neonatal blood spots reveal distinct phenotypes of pediatric acute lymphoblastic leukemia and potential effects of early-life nutrition

Mass Spectrometry-Based Methodologies for Targeted and Untargeted Identification of Protein Covalent Adducts (Adductomics): Current Status and Challenges

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