systeembiologie_metabolomics_09_10b

systeembiologie_metabolomics_09_10b - Introduction What is...

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 5/31/10 Introduction What is metabolomics? The analysis of the full suite of metabolites in a living system What information affords metabolomics? In addition to the proteome, the metabolome level is much closer to the physiology of a living system than the transcriptome . Together, the proteome and metabolome might unravel physiological changes upon a treatment or show the pleiotropic effects on physiology of the altered expression of a gene
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 5/31/10 Approaches in metabolomics
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 5/31/10 Metabolomics Metabolite target analysis : only one metabolite, e.g.  substrate or product of an enzyme Targeted metabolite profiling : only a certain class of  compounds, e.g. phenolics Metabolomics : semi-quantification of as much  metabolites as possible Metabolic fingerprinting : for high-throughput  screening
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 5/31/10 Click to edit Master subtitle style Metabolic fingerprinting NMR IR MS
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 5/31/10 Metabolic fingerprinting Classification of biological samples based on the global metabolome profile as obtained by high-throughput spectroscopic methods. Usually done by direct infusion methods without any chromatography step. Clusters of samples with similar metabolome profiles might be obtained by unsupervised statistical methods, e.g. PCA, or by supervised methods, e.g. DFA. The latter methods incorporate prior knowledge
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 5/31/10 Metabolic fingerprinting: NMR 1. Nuclei have a spin (like electrons) and, thus, a spin angular momentum and a magnetic moment. 2. In the case of 1H, the allowed spin states are +1/2 and -1/2. 3. The nuclei can either align with (+1/2) or oppose (-1/2) the applied magnetic field, Bo (from the NMR instrument).
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 5/31/10 Metabolic fingerprinting: NMR 1. When the nuclei absorb a radiofrequency pulse (a specific energy is absorbed since the spin states are quantized!), the spin flips - resonance. 2. When the pulse is over, the spin relaxes back to its original state. The spin releases the energy that it had originally absorbed – this is the energy that is measured. This happens to all 1H nuclei in the sample, but not all 1H nuclei are the same. 1H nuclei near an electronegative group will be deshielded and feel a stronger Bo. Chemical shift
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 5/31/10 Metabolic fingerprinting: NMR 5.0 0.0 1.0 2.0 3.0 4.0 6.0 7.0 8.0 9.0 10.0 11.0 12.0 Chemical Shift, δ (ppm) CO 2 H C H O Ar H C=C H X-C H O-C H C C H O- H N- H COC H C H , C H 2 C H 3 Gives information on the abundance of the different classes of molecules, such as amines, carbohydrates, phenolics, organic acids,… Some of the most abundant metabolites might be associated with a
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 5/31/10 Metabolic fingerprinting: IR As with NMR, this approach affords information on the abundances of groups of metabolites rather than individual metabolites
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 5/31/10 Metabolic fingerprinting: MS Measures the mass of an ion moving in an electromagnetic field Ionization in the positive mode occurs by uptake of a cation or loss of an electron Ionization in the negative mode occurs by loss of a proton or uptake of a anion Ionisation Chamber Ion Accelerato r Analyser Detector V B r q m 2
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systeembiologie_metabolomics_09_10b - Introduction What is...

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