Cofactor depletion

There is no lag time for inactivation the presence of exogenous nucleophiles has no effect on the inactivation rate following inactivation, a second, equal addition of enzyme results in the same rate of inactivation as the first addition in the absence of inactivator and cofactor depletion. 

The biotransformation of paracetamol (acetaminophen) at therapeutic concentrations in man can be seen in Figure 1. The major route of metabolism is via conjugation of the phenolic -OH group. The hepatic sulphotransferases have a high affinity for paracetamol (acetaminophen) but are rapidly saturated (either due to cofactor depletion or limited sulphotransferase availability), as were the uridine diphosphate glucuronosyl transferases (UDPGTs) which... 

Cofactor depletion Capacity of cell metabolism becomes limiting at higher oxygenase expression levels and/or activities Coexpression of suitable enzymes for cofactor regeneration... 

Another potential complication can occur if the responsiveness of the receptor system changes temporally. This can happen if the receptor (or host system, or both) demonstrates desensitization (tachyphylaxis) to drug stimulation (see Chapter 2). There are numerous systems where constant stimulation with a drug does not lead to a constant steady-state response. Rather, a fade of the response occurs. This can be due to depletion of a cofactor in the system producing the cellular response or a conformational change in the receptor protein. Such phenomena protect against overactive stimulation of... 

Tyrosine is converted to dopa by the cytoplasmic enzyme tyrosine hydroxylase. This is the rate-limiting step 5 x 10 M) in DA synthesis, it requires molecular O2 and Fe + as well as tetrahydropterine (BH-4) cofactor and is substrate-specific. It can be inhibited by a-methyl-p-tyrosine, which depletes the brain of both DA and NA and it is particularly important for the maintenance of DA synthesis. Since the levels of tyrosine are above the for tyrosine hydroxylase the enzyme is normally saturated and so it is not possible to increase DA levels by giving tyrosine. 

Methotrexate is a folic acid analogue. Its mechanism of action is based on the inhibition of dihydrofolate reductase. Inhibition of dihydrofolate reductase leads to depletion of the tetrahydrofolate cofactors that are required for the synthesis of purines and thymidylate (see Fig. 2). Enzymes that are required for purine and thymidylate synthesis are also directly inhibited by the polyglutamates of methotrexate which accumulate with dihydrofolate reductase inhibition. The mechanisms that can cause resistance include decreased transport of methotrexate into the tumor cells, a decreased affinity of the antifolate for dihydrofolate reductase, increased concentrations of intracellular dihydrofolate reductase and decreased thymidylate synthetase activity. 

Mechanism of Action A trace element and essential component in the formation of Hgb. Necessary for effective erythropoiesis and transport and utilization of oxygen. Serves as cofactor of several essential enzymes. Therapeutic Effect Replenishes Hgb and depleted iron stores. 

At blood concentrations below 100 mg/dL (22 mmol/L), the MEOS system, which has a relatively high Km for alcohol, contributes little to the metabolism of ethanol. However, when large amounts of ethanol are consumed, the alcohol dehydrogenase system becomes saturated owing to depletion of the required cofactor, NAD+. As the concentration of... 

Oxygen is normally readily available to all reasonably well-perfused tissues, but deep inside organs such as the liver, especially the centrilobular area (see chap. 6), there will be a reduction in the oxygen concentration. This is clearly important when both oxidative and reductive pathways are available for a particular substrate. Therefore, as conditions in a particular tissue become more anaerobic, reductive pathways will become more important. This is well illustrated by the metabolism of halo thane where, in the rat, hypoxia will increase reductive metabolism and hepa to toxicity (see chap. 7). Glutathione is an extremely important cofactor, involved in both protection and conjugation. It may be depleted by both of these processes, or under certain circumstances, such as hereditary glucose-6-phosphate deficiency in man, supply may be reduced (see chap. 5). This will clearly influence toxicity, and there are a number of examples discussed in chapter 7 in which it is important. 

Of course, one particularly important consideration is dose, as this can dramatically change the balance as a result of saturation or depletion of cofactors. 

The major secondary events are changes in membrane structure and permeability, changes in the cytoskeleton, mitochondrial damage, depletion of ATP and other cofactors, changes in Ca2+ concentration, DNA damage and poly ADP-ribosylation, lysosomal destabilization, stimulation of apoptosis, and damage to the endoplasmic reticulum. 

Depletion of other cofactors such as UTP, NADH, and NADPH may also be involved in cell injury either directly or indirectly. Thus, the role of NADPH in maintaining reduced GSH levels means that excessive GSH oxidation such as caused by certain quinines, which undergo redox cycling, may in turn cause NADPH depletion (see below). Alternatively, NADPH may be oxidized if it donates electrons to the foreign compound directly. However, NADPH may be regenerated by inter conversion of NAD+ to NADP+. Some quinones such as menadione, l,2-dibromo-3-chloropropane (DBCP), and hydrogen peroxide also cause depletion of NAD, but probably by different mechanisms. Thus, with menadione, the depletion may be the result of... 

Isoniazid reacts with pyridoxal phosphate to form a hydrazone (Fig. 7.42), which is a very potent inhibitor of pyridoxal phosphate kinase. The hydrazone has a much greater affinity for the enzyme (100—lOOOx) than the normal substratepyridoxal. The result of this is a depletion of tissue pyridoxal phosphate. This cofactor is of importance particularly in nervous tissue for reactions involving decarboxylation and transamination. The decarboxylation reactions are principally affected however, with the result that transamination reactions assume a greater importance. 

Similarities Both galactosamine and ethionine are toxic as a result of interference with intermediary metabolism. Galactosamine depletes UTP, ethionine depletes ATP. Both form derivatives that effectively trap the cofactor precursor (uridine and adenine, respectively). Both may cause cirrhosis and tumors after repeated dosing. 

Oxaloacetate is formed in the last step of the citric acid cycle by the NAD+-dependent oxidation of L-malate. Can a net synthesis of oxaloacetate from acetyl-CoA occur using only the enzymes and cofactors of the citric acid cycle, without depleting the intermediates of the cycle Explain. How is oxaloacetate that is lost from the cycle (to biosynthetic reactions) replenished ... 

Cell injury can be initiated by a number of mechanisms, such as inhibition of enzymes, depletion of cofactors or metabolites, depletion of energy (ATP) stores, interaction with receptors, and alteration of cell membranes. In recent years attention has focused on the role of biotransformation of chemicals to highly reactive metabolites that initiate cellular toxicity. Many compounds, including clinically useful drugs, can cause cellular damage through metabolic activation of the chemical to highly reactive compounds, such as free radicals, carbenes, and nitrenes (Chapters 7 and 8). 

Protein binding DNA binding SH oxidation Depletion of cofactors Lipid peroxidation... 

Type II deiodinase activity is low in unsupplemented tissue homogenates but is stimulated by DTT [71-74,82,83] and to a lesser extent also by GSH [72]. The DTT concentrations required for maximal enzyme stimulation in the CNS and pituitary seem higher than in BAT and also than those necessary for the type I deiodinase in liver and kidney. Kinetic analysis of the deiodination of varying substrate (T4, rT3) concentrations at different cofactor (DTT) levels have indicated a sequential reaction mechanism for the type II deiodinase [73,82,83]. This is very suggestive of the formation of a ternary enzyme-substrate-cofactor complex in the catalytic process [82], The physiological cofactor of the type II deiodinase has not been identified but it has been observed that GSH depletion with diamide or diethylmaleate impairs T4 to T3 conversion in GH3 pituitary tumor cells [93]. 

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