Dear Solomon,

Plant cytochromes P450 are involved in a wide range of biosynthetic reactions, leading to various fatty acid conjugates, plant hormones, defensive compounds, or medically important drugs. Terpenoids, which represent the largest class of characterized natural plant compounds, are often substrates for plant CYPs.

The analytical chemistry methods are superb for determining structures of products of cytochrome P450 (CYP) reactions. In particular, NMR and mass spectrometry are both sensitive and have high resolving power. Therefore, you can choose a substrate such as terpenoids and monitor its enzymatic metabolism by CYP using H-NMR, LC-MS or any other analytical method. Using the same instruments you can determine the rate of the reactions as well.

For a recent review on this topic, please see the article entitled " Experimental Approaches to Analysis of Reactions of Cytochrome P450 Enzymes" contained in the following link:

http://onlinelibrary.wiley.com/doi/10.1002/9783527673261.ch08/summary

If you are looking on data concerning all available cytochromes, please see the following link which contains a review article entitled "SuperCYP: a comprehensive database on Cytochrome P450 enzymes including a tool for analysis of CYP-drug interactions ":

Nucleic Acids Res. 2010 Jan; 38(Database issue): D237–D243.

Published online 2009 Nov 24. doi:  10.1093/nar/gkp970

PMCID: PMC2808967

SuperCYP: a comprehensive database on Cytochrome P450 enzymes including a tool for analysis of CYP-drug interactions

Saskia Preissner,1 Katharina Kroll,1 Mathias Dunkel,1 Christian Senger,2 Gady Goldsobel,1 Daniel Kuzman,1 Stefan Guenther,1 Rainer Winnenburg,3 Michael Schroeder,3 and Robert Preissner1,*

Author information ► Article notes ► Copyright and License information ►

This article has been cited by other articles in PMC.

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ABSTRACT

Much of the information on the Cytochrome P450 enzymes (CYPs) is spread across literature and the internet. Aggregating knowledge about CYPs into one database makes the search more efficient. Text mining on 57 CYPs and drugs led to a mass of papers, which were screened manually for facts about metabolism, SNPs and their effects on drug degradation. Information was put into a database, which enables the user not only to look up a particular CYP and all metabolized drugs, but also to check tolerability of drug-cocktails and to find alternative combinations, to use metabolic pathways more efficiently. The SuperCYP database contains 1170 drugs with more than 3800 interactions including references. Approximately 2000 SNPs and mutations are listed and ordered according to their effect on expression and/or activity. SuperCYP (http://bioinformatics.charite.de/supercyp) is a comprehensive resource focused on CYPs and drug metabolism. Homology-modeled structures of the CYPs can be downloaded in PDB format and related drugs are available as MOL-files. Within the resource, CYPs can be aligned with each other, drug-cocktails can be ‘mixed’, SNPs, protein point mutations, and their effects can be viewed and corresponding PubMed IDs are given. SuperCYP is meant to be a platform and a starting point for scientists and health professionals for furthering their research.

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2808967/

Hoping this will be helpful,

Rafik

Hi Solomon,

as being long-time involved in PAH effects on mammalian monooxygenases (P450s), your project sounds interesting, in terms of how it might be applicable to earthworms. Especially because the worms experience hypertonic conditions, but less is known about measures employed when faced with hypotonic stress. They possess gene which encodes flavin-containing monooxygenase-4 (FMO-4). FMO-4 is expressed prominently in hypodermis, duct and pore cells but is excluded from the excretory cell. Thus, FMO-4 plays a crucial osmoregulatory role by promoting clearance of excess water that enters during hypotonicity. Most likely, FMO-4 acts via classic heteroatom monooxygenation in an endogenous substrate and the resulting product acts to attenuate water accumulation under hypoosmotic conditions.

Although the specific catalytic activity underlying the osmoregulatory role of FMO-4 has yet to be determined it would be theoretically possible to screen a series of candidate substrates with heterologously expressed enzyme. An alternative and nonbiased approach would be to employ a metabonomics-LC-MSbased system to interrogate C. elegans extracts following incubation with recombinant FMO-4. This approach has been used successfully to identify endogenous ligands for a number of enzymes. By combining the approach with 18O2 labeling the method has also been applied to deorphanize human cytochrome P450 enzymes – a strategy that could also be applied to identify endogenous FMO-4 ligand(s). The increasing number of reports describing FMO involvement in endogenous activities, including atherogenesis and cholesterol metabolism, illustrate these monooxygenases also have, perhaps in many cases, specific biological roles.

Best wishes

Ilya