MRC National Institute for Medical Research, London
Science for Health
Luiz Pedro de Carvalho group project:
Functional genomics of mycobacterial enzymes
Genomics and proteomics continue to deliver a wealth of information about genes and their putative proteins, but experimental functional validation remains a great challenge. For instance, hundreds of genes from Mycobacterium tuberculosis (Mtb) are predicted to encode enzymes that are involved in biosynthesis and degradation of cofactors, amino acids, sugars and lipids, but very few of these have had their function revealed and validated. Approximately 35% of the Mtb genome is composed of essential genes, which could be potential targets for pharmacological intervention, if their function was known.
Scheme of the activity-based metabolomics profiling method for functional genomics
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The left panel illustrates the process used to prepare our substrate library (small molecule extract or metabolome). The middle panel shows the set up for the enzymatic reaction and control. The right panel illustrates the separation, identification and quantitation obtained by accurate mass LC-MS and the next steps that define reaction (based on a substrate and product pair) and the putative pathway where it belongs.
I plan to expand our activity-based metabolomics profiling (ABMP) and test its applicability to different enzyme classes, beginning with putative and uncharacterized S-adenosylmethionine-dependent methyltransferases and NAD(P)+/NAD(P)H-utilizing oxidoreductases. This method uses the metabolome of the cell as a highly dense, chemically diverse yet physiologically relevant small molecule library (left panel), recombinant enzyme as a catalyst (middle panel), and accurate mass LC-MS as analytical technique (right panel). Validation of the observed chemistries will be performed using standard genetic, biochemical and metabolomics approaches. Using this approach we will reveal new enzymatic functions, discover new small molecules and chemistries, and map uncharacterized biochemical pathways, some of which will be essential during Mtb infection.
de Carvalho group
Recent publications
Research projects
- Functional genomics of mycobacterial enzymes
- Rational antibiotic discovery and systems pharmacology
- Metabolomics-level physiology studies
de Carvalho biography
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