Dataset Information

OxiTMT - Quantitative analysis of the cysteine redoxome by iodoacetyl tandem mass tags

ABSTRACT: Developpment on a new proteomics analytical tool for the the study of reversible cysteine oxidation by isobaric labeling.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Escherichia Coli

SUBMITTER: Shakir Shakir

LAB HEAD: Joelle VINH

PROVIDER: PXD005164 | Pride | 2018-07-27

REPOSITORIES: Pride

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Quantitative analysis of the cysteine redoxome by iodoacetyl tandem mass tags.

Shakir Shakir S Vinh Joelle J Chiappetta Giovanni G

Analytical and bioanalytical chemistry 20170407 15

The redox conditions that reign inside a cell have a determining effect on a number of biological processes. Reactive oxygen species (ROS) and reactive nitrogen species (RNS) are key redox players and have been linked to a number of pathologies. They have also been shown to play an important regulating role in cell signaling events. On the proteome level, thiol groups of cysteinyl side chains constitute the major targets of ROS and RNS. A number of analytical techniques based on mass spectrometr ...[more]

PMID: 28389918

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Project description:Milk oligosaccharides (OS) are a key factor that influences the infant gut microbial composition, and their importance in promoting healthy infant development and disease prevention is becoming increasingly apparent. Investigating the structures, properties, and sources of these compounds requires a host of complementary analytical techniques. Relative compound quantification by mass spectral analysis of isobarically labeled samples is a relatively new technique that has been used mainly in the proteomics field. Glycomics applications have so far focused on analysis of protein-linked glycans, while analysis of free milk OS has previously been conducted only on analytical standards. In this paper, we extend the use of isobaric glycan tags to the analysis of bovine milk OS by presenting a method for separation of labeled OS on a porous graphitized carbon liquid chromatographic column with subsequent analysis by quadrupole time-of-flight tandem mass spectrometry. Abundances for 15 OS extracted from mature bovine milk were measured, with replicate injections providing coefficients of variation below 15% for most OS. Isobaric labeling improved ionization efficiency for low-abundance, high-molecular weight fucosylated OS, which are known to exist in bovine milk but have been only sporadically reported in the literature. We compared the abundances of four fucosylated OS in milk from Holstein and Jersey cattle and found that three of the compounds were more abundant in Jersey milk, which is in general agreement with a previous study. This novel method represents an advancement in our ability to characterize milk OS and provides the advantages associated with isobaric labeling, including reduced instrumental analysis time and increased analyte ionization efficiency. This improved ability to measure differences in bioactive OS abundances in large datasets will facilitate exploration of OS from all food sources for the purpose of developing health-guiding products for infants, immune-compromised elderly, and the population at large.

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	LFpeptides.txt	Txt
	LFproteinGroups.txt	Txt
	QE150724_1343_i_ssh.raw	Raw
	QE150724_1344_i_ssh.raw	Raw
	QE150724_1345_i_ssh.raw	Raw