Cytochrome P450

By Jennifer McDowall
The elimination of foreign compounds (xenobiotics) such as drugs and toxins
from the body is an essential process designed to protect against potential toxicity from
the foods we eat. The food broken down in the stomach is absorbed by the small
intestine and then ferried directly to the liver via the portal vein (see figure below). This
allows the liver time to detoxify compounds before they are distributed through the
circulatory system. In the liver, there are two main types of metabolism that deal with
xenobiotics, and a third that deals with their transport.

· Phase I metabolism results in small chemical changes that make a compound

more hydrophilic, so it can be effectively eliminated by the kidneys. These reactions
usually involve either adding or unmasking a hydroxyl group, or some other hydrophilic
group such as an amine or sulphydryl group, and usually involve hydrolysis, oxidation or
reduction mechanisms. Cytochrome P450 enzymes are responsible for most phase I
reactions.

· Phase II metabolism takes place if phase I is insufficient to clear a compound

from circulation, or if phase I generates a reactive metabolite. These reactions usually
involve adding a large polar group (conjugation reaction), such as glucuronide, to further
increase the compound’s solubility. Often, the functional groups generated in phase I
reactions are required for attachment of the phase II polar groups (though in some
cases phase II reactions can occur on their own). Transferase enzymes are
responsible for most phase II reactions, e.g. uridine diphosphoglucuronosyl transferase
(UGT), N-acetyl transferase (NAT), glutathione S-transferase (GST), and
sulphotransferase (ST).

· Phase III involves drug transporters, which influence the effect, absorption,
distribution and elimination of a drug. Drug transporters move drugs across cellular
barriers, and as such can target sites of accumulation. They are located in epithelial
and endothelial cells of the liver, gastrointestinal tract, kidney, blood-brain barrier and
other organs.

Cytochrome P450 enzymes are the most important enzymes in Phase I metabolism in
mammals, and are primarily responsible for the metabolism (degradation and
elimination) of drugs.
 Liver portal system: food is digested in the stomach and small intestine, then
absorbed in the small intestine, where it is transported to the liver for detoxification
before being distributed.

Cytochrome P450, Detox Enzymes

Cytochrome P450 (CYP) enzymes are a superfamily of mono-oxygenases that
are found in all kingdoms of life, and which show extraordinary diversity in their reaction
chemistry. In mammals, these enzymes are found primarily in the membranes of the
endoplasmic reticulum (microsomes) within liver cells (hepatocytes), as well as many
other cell types. These enzymes use haem iron to oxidise molecules, often making
them more water-soluble for clearance. They achieve this by either adding or
unmasking a polar group. In general, the reaction catalysed by these enzymes can be
summarised as:

R-H + O2 + 2e- + 2H+ à R-OH + H2O

Where R-H is the substrate and R-OH is the oxygenated substrate. The oxygen is
bound to the haem in the core of the CYP enzyme. Protons (H+) are usually delivered
from the cofactor NADH or NADPH through specific amino acids in the CYP enzyme,
which relay the protons to the active site, where they are essential for a reductive
splitting of the oxygen so a single atom can be added to the substrate. CYP enzymes
can receive electrons from a range of different redox partner enzymes.
Mammalian enzymes

Mammalian CYP enzymes can oxidise both xenobiotics and endogenous

compounds, and are important for detoxification of foreign substances, as well as for
controlling the level of endogenous compounds, such as hormone synthesis and
breakdown, cholesterol synthesis and vitamin D metabolism. CYP enzymes are also
involved in vascular autoregulation, especially in the brain, and are vital to the formation
of cholesterol, steroids and arachidonic acid metabolites. They can also clear the body
of metabolic products such as bilirubin, which arises from the breakdown of
haemoglobin. There is a high concentration of CYP proteins in the liver, but these
enzymes are also found throughout the body, where they often have specialised roles.

Plant enzymes

Plant CYP enzymes are important for the biosynthesis of several compounds,
such as hormones, defensive compounds, and fatty acid conjugates.

Bacterial enzymes

Bacterial CYP enzymes are important for several metabolic processes, such as
the camphor-hydroxylating catalytic cycle in P. putida, and for the biosynthesis of the
antibiotic erythromycin in S. erythraea.