Metabolomic Analysis of <i>Schizosaccharomyces pombe</i>: Sample Preparation, Detection, and Data Interpretation

During human fasting, metabolic markers, including butyrates, carnitines, and branched-chain amino acids, are upregulated for energy substitution through gluconeogenesis and use of stored lipids. We performed non-targeted, accurate semiquantitative metabolomic analysis of human whole blood, plasma, and red blood cells during 34–58 hr fasting of four volunteers. During this period, 44 of ~130 metabolites increased 1.5~60-fold. Consistently fourteen were previously reported. However, we identified another 30 elevated metabolites, implicating hitherto unrecognized metabolic mechanisms induced by fasting. Metabolites in pentose phosphate pathway are abundant, probably due to demand for antioxidants, NADPH, gluconeogenesis and anabolic metabolism. Global increases of TCA cycle-related compounds reflect enhanced mitochondrial activity in tissues during fasting. Enhanced purine/pyrimidine metabolites support RNA/protein synthesis and transcriptional reprogramming, which is promoted also by some fasting-related metabolites, possibly via epigenetic modulations. Thus diverse, pronounced metabolite increases result from greatly activated catabolism and anabolism stimulated by fasting. Anti-oxidation may be a principal response to fasting.

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“…Metabolite standards were purchased from commercial sources (Table S3). Other reagents have been previously described 6,7,43 .…”

Section: Methodsmentioning

confidence: 99%

Diverse metabolic reactions activated during 58-hr fasting are revealed by non-targeted metabolomic analysis of human blood

Teruya

Chaleckis

Takada

et al. 2019

Sci Rep

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“…Metabolite standards were purchased from commercial sources (Table S3). Other reagents have been previously described 6,7,43 . Hospital and by the Human Subjects Research Review Committee of the Okinawa Institute of Science and Technology Graduate University (OIST).…”

Section: Methodsmentioning

confidence: 99%

Diverse Metabolic Reactions Activated During 58-hr Fasting are Revealed by Non-Targeted Metabolomic Analysis of Human Blood

et al. 2018

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During human fasting, metabolic markers, including butyrates, carnitines, and branched-chain amino acids, are upregulated for energy substitution through gluconeogenesis and use of stored lipids. We performed non-targeted, accurate semiquantitative metabolomic analysis of human whole blood, plasma, and red blood cells during 34-58 hr fasting of four volunteers. During this period, 44 of ~130 metabolites increased 1.5~60-fold. Consistently fourteen were previously reported. However, we identified another 30 elevated metabolites, implicating hitherto unrecognized metabolic mechanisms induced by fasting. Metabolites in pentose phosphate pathway are abundant, probably due to demand for antioxidants, NADPH, gluconeogenesis and anabolic metabolism. Global increases of TCA cyclerelated compounds reflect enhanced mitochondrial activity in tissues during fasting. Enhanced purine/ pyrimidine metabolites support RNA/protein synthesis and transcriptional reprogramming, which is promoted also by some fasting-related metabolites, possibly via epigenetic modulations. Thus diverse, pronounced metabolite increases result from greatly activated catabolism and anabolism stimulated by fasting. Anti-oxidation may be a principal response to fasting.

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“…We initially investigated metabolomics of a simple model eukaryote, fission yeast (Schizosaccharomyces pombe) using yeast genetic technology to identify and determine metabolite profiles in wild type and mutant cell extracts (Pluskal et al, 2010b) (Pluskal and Yanagida, 2016a) (Pluskal and Yanagida, 2016b). By adapting the software MZmine 2 for metabolite identification (Pluskal et al, 2010a) (Pluskal et al, 2012), we found that metabolomics of fission yeast and humans are surprisingly similar in regard to metabolite composition (Chaleckis et al, 2014).…”

Section: Discussionmentioning

confidence: 99%

Aging markers in human urine: A comprehensive, non-targeted LC-MS study

Teruya

Goga

Yanagida

2020

Preprint

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Age markers in urine, non-targeted metabolomics, human aging, LC-MS, highly 23 correlated metabolites, leucine, pseudouridine, 24 25 Short title: Aging markers in human urine 26 27 Summary 28Metabolites in human biofluids document individual physiological status. We 29 conducted comprehensive, non-targeted, non-invasive metabolomic analysis of urine 30 from 27 healthy human subjects, comprising 13 youths (30±3 yr) and 14 seniors 31 (76±4 yr). Quantitative analysis of 99 metabolites revealed 55 that were linked to 32 aging, displaying significant differences in abundance between the two groups. 33These include 13 standard amino acids, 5 methylated, 4 acetylated, and 9 other 34 amino acids, 6 nucleosides, nucleobases, and derivatives, 4 sugar derivatives, 5 35 sugar phosphates, 4 carnitines, 2 hydroxybutyrates, 1 choline, and 1 ethanolamine 36 derivative, and glutathione disulfide. Abundances of 53 compounds decreased, while 37 2 increased in elderly people. Many age-linked markers were highly correlated; 42 of 38 55 compounds, showed Pearson's correlation coefficients larger than 0.70. As 39 metabolite profiles of urine and blood are quite different, age-related information 40 in urine components offer yet more valuable insights into aging mechanisms of 41 endocrine system and related organ systems. 42 43Results 71 Collection of urine samples 72Samples of morning urine, immediately after awaking, were collected 73 from healthy volunteer subjects (elderly, 75.8±3.9 yr, and young, 30.6±3.2 yr; 74 Supplementary Table s1 shows gender and BMI) in Onna Village, Okinawa, 75Japan. Precautions taken for sample collection are described in the Materials 76 and Methods. Basic data analytical procedures were similar to those previously 77 described (Chaleckis et al., 2016) (Teruya et al., 2019) (Kameda et al., 2020). 78 79 99 urine metabolites identified 80 Ninety-nine urinary metabolites, about half of which are amino acids and 81 their derivatives, were identified and quantified using liquid chromatography -mass 82 spectrometry (LC -MS) and MZmine 2 (Pluskal et al., 2010a) (Teruya et al., 2019) 83 (Supplemental Table s2). These compounds were subdivided into 12 groups, 84 containing 17 standard amino acids, 12 methylated amino acids, 6 acetylated amino 85 acids and 15 other amino acids, 12 nucleosides, nucleobases, and derivatives, 4 sugar 86 derivatives, 6 sugar phosphates, 3 vitamins and coenzymes (pantothenate, 4-87 aminobenzoate, nicotinamide), 4 choline and ethanolamine derivatives, 8 carnitines, 88 11 organic acids, and 1 antioxidant (oxidized form of glutathione, GSSG). Small amino 89 acids, such as glycine and alanine, were not detected in our analysis due to the 90 mass cutoff (100 m/z) used. Levels of individual compounds, categorized by 91 abundance as H (high, >10 8 ), M (medium, 10 7~1 0 8 ) or L (low, <10 7 ), were estimated 92 based upon mass spectroscopic peak area (Chaleckis et al., 2014) (Chaleckis et al., 93 6 2016). Some compounds varied widely from one individual to the next and are 94 denoted as H-L, H-...

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Metabolomic Analysis of Schizosaccharomyces pombe: Sample Preparation, Detection, and Data Interpretation

Cited by 6 publications

References 59 publications

Diverse metabolic reactions activated during 58-hr fasting are revealed by non-targeted metabolomic analysis of human blood

Diverse metabolic reactions activated during 58-hr fasting are revealed by non-targeted metabolomic analysis of human blood

Diverse Metabolic Reactions Activated During 58-hr Fasting are Revealed by Non-Targeted Metabolomic Analysis of Human Blood

Aging markers in human urine: A comprehensive, non-targeted LC-MS study

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