31 CHAPTER 4 Damage Process and Action of Toxicants 4.1 INTRODUCTION At a sufficiently high concentration, a pollutant can elicit adverse effects on the living processes of an organism. To exert damage to an exposed organism, the pollutant must first enter the host and reach its target site. A complex pathway exists from the time of initial toxicant exposure to manifestation of damage within the organism. This chapter discusses general ways in which environmental pollutants exert their actions on plants, animals, and humans. 4.2 PLANTS 4.2.1 Sources of Pollution For the most part, environmental pollution is an anthropogenic (human-made) problem. The most important source of atmospheric pollution in the United States is motor vehicle transportation. Other major sources include industry, power gener- ation, space heating, and refuse burning. The composition of pollutants from various sources differs markedly, with industry emitting the most diversified pollutants. While carbon monoxide (CO) is the major component of pollution by motor vehicles, sulfur oxides (SO x ) are primary pollutants of industry, power generation, and space heating. In some large cities, such as Los Angeles, accumulation of O 3 , PAN, and other photochemical oxidants constitutes the major atmospheric pollution problem. 4.2.2 Pollutant Uptake Terrestrial plants may be exposed to environmental pollutants in two main ways. One is exposure of foliage to air pollutants; another is uptake of toxicants by roots growing in contaminated soils. Vegetation growing near industrial facilities, such as smelters, aluminum refineries, and coal-burning power plants, may absorb airborne LA4154/frame/C04 Page 31 Wednesday, May 17, 2000 3:52 PM © 2001 by CRC Press LLC 32 ENVIRONMENTAL TOXICOLOGY pollutants into the leaves and become injured. The pollutants may be in gaseous form such as SO 2 , NO 2 , and HF, or in particulate form such as the oxides or salts of metals contained in fly ash (Figure 4.1). To consider the effects of any airborne pollutants on vegetation, the uptake of the pollutants by the plant is critical. While the atmospheric concentration of a pollutant is an important factor, the actual amount that enters the plant is more important. The conductance through the stoma, which regulates the passage of ambient air into the cells, is especially critical. The extent of uptake depends on the chemical and physical properties along the gas-to-liquid diffusion pathway. The flow of a pollutant may be restricted by the leaf’s physical structure or by scavenging chemical reactions occurring in the leaf. Leaf orientation and morphology, including epidermal characteristics, and air movement across the leaf are important determi- nants affecting the initial flux of gases to the leaf surface. Stomatal (Figure 4.2) resistance is a very important factor affecting pollutant uptake. The resistance is determined by stomatal size, number, and anatomical characteristics, and the size of the stomatal aperture. Little or no uptake may occur when the stoma is closed. Stomatal opening is regulated by light, humidity, temper- ature, internal CO 2 content, water and nutrient availability, and K + ions transported into the guard cells. 1 Exposure of roots to toxicants in contaminated soils is another important process whereby toxicant uptake by plants occurs. For example, vegetation growing in soils Figure 4.1 Air pollutants may damage trees in different ways. LA4154/frame/C04 Page 32 Wednesday, May 17, 2000 3:52 PM © 2001 by CRC Press LLC DAMAGE PROCESS AND ACTION OF TOXICANTS 33 of contaminated sites, such as waste sites and areas that have received contaminated sewage sludge, can absorb toxicants from the root. In the contaminated sites, high levels of heavy metals such as Pb and Cd often occur. Metallic ions are more readily released when the soil is acidified by acid deposition (Figure 4.1). 4.2.3 Transport Following uptake, a toxicant may undergo mixing with the surrounding medium and then be transported to various organs and tissues of the plant. Mixing involves a microscopic movement of molecules and is accompanied by compensation of concentration differences. Generally, transport of chemicals in plants occurs pas- sively by diffusion and flux. Diffusion refers to movement across phase boundaries, from a high concentration compartment to a low concentration compartment. Flux, on the other hand, is due to the horizontal movement of the medium. 4.2.4 Plant Injury Besides killing plants, air pollutants adversely affect plants in various ways. Pollution injury is commonly divided into acute or chronic injury. In plants, an acute injury occurs following absorption of sufficient amounts of toxic gas or other forms of toxicants to destroy the tissue. It is often characterized by collapsed marginal or other areas of the leaf with an initial water-soaked appearance. Subsequently, they dry and bleach to an ivory color or become brown or brownish red. Chronic injury, on the other hand, results from absorption of gaseous or other forms of pollutants that are somewhat insufficient to cause acute injury, or it may be caused by uptake of sublethal amounts of toxicants over a long period of time. Chronic injury is manifested by leaf yellowing that may progress slowly through stages of bleaching until most of the chlorophyll and carotenoids are destroyed. Concerning leaf injury caused by atmospheric pollution, the epidermis is the first target as an air pollutant passes through the stomata of the epidermal tissue Figure 4.2 Cross section of intact leaf. The air spaces within a leaf serve as passages for pollutants that may subsequently injure the leaf. Stoma Guard Cells (enlarged) Lower Epidermal Cells Vascular Bundle Spongy Parenchyma Mesophyll Cells Palisade Parenchyma Air Spaces Upper Epidermal Cells LA4154/frame/C04 Page 33 Wednesday, May 17, 2000 3:52 PM © 2001 by CRC Press LLC 34 ENVIRONMENTAL TOXICOLOGY (Figure 4.2). In passing into the intercellular spaces, the pollutant may dissolve in the surface water of the leaf cells, affecting cellular pH. A pollutant might not remain in its original form as it passes into solution. Rather, it might be converted into a form that is more reactive and toxic than the original substance. The formation of reactive free radicals following the initial reaction in the cell is an example. The pollutant, either in its original form or in an altered form, may then react with certain cellular components such as the cytoplasmic membrane or the membranes of the organelles, enzymes, coenzymes or cofactors, and substrates. The pollutant may thus affect cellular metabolism leading to plant injury. Sulfur dioxide-induced changes in ultrastructure of various organelles, such as chloroplasts or mitochondria, can disrupt photosynthesis or cellular energy metabolism. As a pollutant moves from the substomatal regions to the cellular sites of perturbation, it may encounter various obstacles along the pathway. Scavenging reactions between endogenous substances and the pollutant might occur, and the result can affect pollutant toxicity itself. For instance, ascorbate, which occurs widely in plant cells, might react with or neutralize a particular pollutant or a secondary substance formed as the pollutant is metabolized. On the other hand, an oxidant such as O 3 might react with membrane material and induce peroxidation of the lipid components. This can lead to the formation of toxic substances such as aldehydes, ketones, and free radicals. The free radicals, in turn, can attack various cellular components, including proteins, lipids, and nucleic acids, leading to tissue damage. Endogenous antioxidants, such as the ascorbic acid mentioned above, may react with free radicals and alter their toxicity. Cellular enzyme inhibition often occurs when leaves are exposed to various atmospheric pollutants. For instance, fluoride (F) is widely known as a metabolic inhibitor and as such can inhibit a number of enzymes. Often, such enzyme inhibition is attributable to reaction of F – with certain cofactors such as Ca 2+ or Mg 2+ in an enzyme system. Heavy metals such as Pb and Cd may also inhibit enzymes that contain sulfhydryl (–SH) groups at the active sites. On the other hand, SO 2 may oxidize and break apart the sulfur bonds in the critical enzymes of the membrane, thus disrupting cellular function. As noted above, soil acidification increases release of toxic metal ions. These metal ions may directly damage roots through disrupting water and nutrient uptake, resulting in water deficit and nutrient deficiency. Soil acidification can also cause leaching of nutrients, leading to nutrient deficiency and growth disturbance (Figure 4.1). Plants become unhealthy as a result of one or more of the disturbances men- tioned above. Even before visible symptoms are discernible, an exposed plant may be weakened and its growth impaired. In time, visible symptoms such as chlorosis or necrosis may appear, followed by death of the plant. 4.3 MAMMALIAN ORGANISM 4.3.1 Exposure An environmental pollutant may enter an animal or human through a series of pathways. Figure 4.3 shows the general pathway pollutants may pass through during LA4154/frame/C04 Page 34 Wednesday, May 17, 2000 3:52 PM © 2001 by CRC Press LLC DAMAGE PROCESS AND ACTION OF TOXICANTS 35 their presence in a mammalian organism. As mentioned earlier, exposure to a pol- lutant by a host organism constitutes the initial step in the manifestation of toxicity. A mammalian organism may be exposed to pollutants through inhalation, dermal or eye contact, or ingestion. 4.3.2 Uptake The immediate and long-term effects of pollutants are directly related to the mode of entry. The portals of entry for an atmospheric pollutant are the skin, eyes, lungs, and gastrointestinal tract. The hair follicles, sweat glands, and open wounds are possible entry sites where uptake from the skin may occur. Both gaseous and particulate forms of air pollutants can be taken up through the lungs. Uptake of toxicants by the gastrointestinal tract may occur when consumed foods or beverages are contaminated by air pollutants such as Pb and Cd, or sprayed pesticides. For a pollutant to be taken up into the body and finally carried to the cell, it must pass through several biological membranes. These include not only the periph- Figure 4.3 The poisoning process in animals and humans. LA4154/frame/C04 Page 35 Wednesday, May 17, 2000 3:52 PM © 2001 by CRC Press LLC 36 ENVIRONMENTAL TOXICOLOGY eral tissue membranes, but also the capillary and cell membranes. Thus, the nature of the membranes and the chemical and physical properties (e.g., lipophilicity) of the toxicant in question are important factors affecting uptake. The mechanisms by which chemical agents pass through the membrane include: (a) filtration through spaces or pores in membranes; (b) passive diffusion through the spaces or pores, or by dissolving in the lipid material of the membrane; (c) facilitated transport, where a specialized molecule called a “carrier,” a protein, carries a water-soluble substance across the membrane; and (d) active transport, which requires both a “carrier” and energy (Table 4.1). Of the four mechanisms, active transport is the only one in which a toxicant may move against a concentration gradient, i.e., move from a low con- centration compartment to a high concentration compartment. This is the reason for the requirement of energy expenditure in active transport. 4.3.3 Transport Immediately after absorption, a toxicant may be bound to a blood protein (such as a lipoprotein) forming a complex, or it may exist in a free form. Rapid transport throughout the body follows. Transport of a toxicant may occur via the bloodstream or lymphatic system. The toxicant may then be distributed to various body tissues, including those of storage depots and sites of metabolism or biotransformation. 4.3.4 Storage A toxicant may be stored in the liver, lungs, kidneys, bone, adipose tissue, and other sites. These storage depots may or may not be the sites of the toxic action. A toxicant may be stored in a depot temporarily and then removed and translocated again. Similarly, a toxicant or its metabolite may be transported to a storage site and remain there for a long period of time or permanently. Excretion of the toxicant following a temporary storage in a storage depot can also occur. 4.3.5 Metabolism The metabolism of toxicants may occur at portals of entry or in such organs as skin, lungs, liver, kidney, and the gastrointestinal tract. The liver plays a central role in the metabolism of environmental toxicants or xenobiotics. Metabolism of xenobi- Table 4.1 Four Basic Types of Absorption Processes Process Energy Need Carrier Concentration Gradient Passive no no high → low Facilitated no yes high → low Active yes yes high → low low → high Phagocytosis/Pinocytosis a yes no NA a Phagocytosis is involved in invagination of a solid particle, whereas pinocytosis involves liquid. LA4154/frame/C04 Page 36 Wednesday, May 17, 2000 3:52 PM © 2001 by CRC Press LLC DAMAGE PROCESS AND ACTION OF TOXICANTS 37 otics is often referred to as biotransformation. A rich supply of nonspecific enzymes exists in the liver, enabling it to metabolize a broad spectrum of organic compounds. The reactions included in biotransformation are classified into two pathways, Phases I and II. Phase I reactions are often divided into three main categories, i.e., oxidation, reduction, and hydrolysis. They are characterized by introduction of a reactive polar group into the xenobiotic, forming a primary metabolite. Phase II reactions, on the other hand, involve conjugation reactions in which an endogenous substance combines with the primary metabolite, forming a complex secondary metab- olite. The resultant secondary metabolite is more water soluble, and thus more readily excretable than the original toxicant. While many xenobiotics are detoxified through these reactions, others may be converted to more active and more toxic compounds. We will discuss biotransfor- mation in more detail in the next chapter. 4.3.6 Excretion The final step involved in the action of a toxicant is its excretion from the body. Excretion may occur through the lungs, kidneys, or intestinal tract. A toxicant may be excreted in its original form or as its metabolite(s), depending on the chemical properties. Excretion is the most permanent means whereby toxic substances are removed from the body. 4.4 MECHANISM OF ACTION The toxic action of pollutants involves compounds with intrinsic toxicity or activated metabolites. These interact with cellular components at their site of action to initiate toxic effects, which may occur anywhere in the body. The consequence of such action may be reflected in changes in physiological and biochemical pro- cesses in the exposed organism. Such changes may be manifested in different ways. Examples are impaired oxidative metabolism and the central nervous system (CNS), injury to the reproductive system, or interaction with nucleic acids leading to car- cinogenesis. The action of a toxicant may be terminated by storage, biotransforma- tion, or excretion. The mechanism involved in the manifestation of toxicant-induced toxicity is complex and much remains to be elucidated. Nevertheless, several representative examples are given here. Generally, a toxicant may cause an adverse effect on living organisms by (a) disruption or destruction of cellular structure; (b) direct chemical combination with a cell constituent; (c) inhibition of enzymes; (d) initiation of a secondary action; (e) free radical-mediated reactions; and (f) disruption of repro- ductive function. These are examined in the following pages. 4.4.1 Disruption or Destruction of Cellular Structure A toxicant may induce an injurious effect on plant or animal tissues by disrupting or destroying the cellular structure. For instance, atmospheric pollutants such as LA4154/frame/C04 Page 37 Wednesday, May 17, 2000 3:52 PM © 2001 by CRC Press LLC 38 ENVIRONMENTAL TOXICOLOGY SO 2 , NO 2 , and O 3 are phytotoxic — they can cause plant injury. Sensitive plants exposed to any of these pollutants at sufficiently high concentrations may exhibit structural damage resulting from cellular destruction. Research has shown that low concentrations of SO 2 can injure epidermal and guard cells, leading to enhanced stomatal conductance and greater entry of the pollutant into leaves. 2 Similarly, after entry into the substomatal cavity of the plant leaf, O 3 , or the free radicals produced from it, may react with protein or lipid membrane components, disrupting the cellular structure of the leaf. 3,4 In animals and humans, sufficient quantities of inhaled NO 2 and sulfuric acid mists can damage surface layers of the respiratory system. Similarly, high levels of O 3 can induce peroxidation of polyunsaturated fatty acids in the cell membrane, thus disrupting the membrane structure. 5 4.4.2 Chemical Combination with a Cell Constituent A pollutant may combine with a cell constituent, forming a complex and dis- rupting cellular metabolism. For example, a number of toxicants or their metabolites are capable of binding to DNA to form DNA adducts. Formation of such adducts results in structural changes in DNA and disrupts its function, and may lead to carcinogenesis. For instance, benzo(a)pyrene, one of the many polycyclic aromatic hydrocarbons (PAHs), can be converted to its epoxide in the body. The resultant epoxide can in turn react with guanine on the DNA molecule, forming a guanine adduct. Many alkylating agents are metabolized to reactive alkyl radicals capable of adduct formation as well. These will be discussed in more detail in Chapter 15. Certain metallic cations can interact with the anionic phosphate groups of poly- nucleotides. They can also bind to polynucleotides at various specific molecular sites, particularly purines and thymine. Such metallic cations can, therefore, inhibit DNA replication and RNA synthesis and cause nucleotide mispairing in polynucleotides. An anatomical feature of chronic intoxication of Pb in humans and in various animals is the presence of characteristic intranuclear inclusions in proximal tubular epithelial cells of the kidneys. These inclusions appear to be formed from Pb and soluble proteins. 6 By tying up cellular proteins, Pb can depress or destroy their function. Another example is the binding of CO to hemoglobin. After it is inhaled and is present in the blood, CO readily reacts with hemoglobin (Hb) to form carboxyhe- moglobin (COHb): CO + Hb → COHb (4.1) The presence of a large amount of COHb in the blood disrupts the vital CO 2 –O 2 exchange system between the lungs and other body tissues. Interference with the functioning of hemoglobin by COHb accumulation is detrimental to health and can lead to death. 4.4.3 Effect on Enzymes The most distinguishing feature of reactions that occur in a living cell is the participation of enzymes as biological catalysts. Almost all enzymes are proteins LA4154/frame/C04 Page 38 Wednesday, May 17, 2000 3:52 PM © 2001 by CRC Press LLC DAMAGE PROCESS AND ACTION OF TOXICANTS 39 and have globular structure, and many of them carry out their catalytic function by relying solely on their protein structure. Many others require nonprotein components called cofactors , which may be metal ions or organic molecules referred to as coenzymes . Metal ions capable of acting as cofactors include K + , Na + , Cu 2+ , Fe 2+ , Mg 2+ , Mn 2+ , Ca 2+ , and Zn 2+ ions. In addition, there are several nonmetal elements that have a function similar to the metal ions in an enzyme system. Table 4.2 shows several metal ions that some enzymes require for their action. Many coenzymes are vitamins or contain vitamins as part of their structure. Usually, a coenzyme is firmly bound to its enzyme protein, and it is difficult to separate the two. Such tightly bound coenzymes are referred to as prosthetic groups of the enzyme. The catalytically active complex of protein and prosthetic group is called the holoenzyme , while the protein without the prosthetic group is called the apoenzyme , which is catalytically inactive. Enzyme + prosthetic group → Protein–prosthetic group (4.2) ( Apoenzyme )( Holoenzyme ) Coenzymes are especially important in animal and human nutrition because, as previously mentioned, most of them are vitamins or are substances produced from vitamins. For instance, following ingestion, niacin, a B vitamin, is converted to nicotinamide adenine dinucleotide (NADH) or nicotinamide adenine dinucleotide phosphate (NADPH). Both NADH and NADPH are important coenzymes in cellular metabolism. There are several ways in which toxicants can inhibit enzymes, leading to disruption of metabolic pathways. Some examples are given below: (a) A toxicant may inhibit an enzyme by inactivating the cofactor . As mentioned above, some cofactors are metallic ions, and they provide electrophilic centers in the active site, facilitating catalytic reactions. For instance, fluoride (F) has been shown to inhibit α -amylase, an enzyme responsible for the breakdown of starch into maltose and eventually glucose. α -Amylase is known to require Ca 2+ for its stability as well as its catalytic action. 7,8 In the presence of F – ions, α -amylase activity is depressed. 9,10 The mechanism involved in the inhibition appears to be through removal of the Ca 2+ cofactor by F – ions. 10 Evidence supporting this observation was obtained when a crude enzyme extract from seedlings exposed to 5 m M NaF for 3 days and incubated with CaCl 2 exhibited a higher α -amylase activity than the control assay mixture containing no added CaCl 2 (Figure 4.4). Table 4.2 Metallic Ions and Some Enzymes that Require Them Metallic Ion Enzyme Ca 2+ Lipase, α -amylase Cu 2+ Cytochrome oxidase Fe 2+ or Fe 3+ Catalase; cytochrome oxidase; peroxidase K + Pyruvate kinase (also requires Mg 2+ ) Mg 2+ Hexokinase, ATPase, enolase Zn 2+ Carbonic anhydrase; DNA polymerase LA4154/frame/C04 Page 39 Wednesday, May 17, 2000 3:52 PM © 2001 by CRC Press LLC 40 ENVIRONMENTAL TOXICOLOGY Fluoride is also known to inhibit enolase, an enzyme involved in glycolysis. Enolase requires Mg 2+ as cofactor. The F-induced inhibition of the enzyme is more marked in the presence of phosphate. It is generally assumed that the mechanism involved in the inhibition is due to inactivation of the Mg through formation of a magnesium-fluoro-phosphate. (b) A toxicant may exert its toxicity by competing with the cofactor of an enzyme , thereby inhibiting its activity . Many enzymes require Zn 2+ ions as a cofactor for optimal activity (Table 4.2). Cadmium (Cd 2+ ), which is chemically similar to Zn 2+ , exhibits inhibition of these enzymes by competing with the Zn 2+ cofactor. Also, beryllium (Be) is known to inhibit certain enzymes that require Mg 2+ for a similar reason. (c) A toxicant may bind to the active site of an enzyme, depressing its activity . For instance, a thiol or sulfhydryl (SH) group on a protein enzyme is often the active site for the enzyme to perform its catalytic action. A heavy metal, such as Pb, Cd, or Hg, after absorption into the body may attach itself to the SH group, forming a covalent bond with the sulfur atom (Equation 4.3). With the active site blocked, the activity of the enzyme may be depressed or lost. 2 Enz -SH + Pb 2+ → Enz -S- Pb -S- Enz + 2H + (4.3) It has been shown that transaminases and δ -aminolevulinate dehydratase both have SH groups as their active sites. They are, therefore, susceptible to Pb inhibition. Figure 4.4 Effect of Ca on α -amylase activity in mung bean seedlings exposed to NaF. Enzyme extracts were prepared from seedlings exposed to 5.0 m M NaF for 24 h. The enzyme assay mixture contained Tris-buffer (pH 7.0), 0.2% starch solution, and water (control) or 5 m M CaCl 2 , and the mixture was incubated for a total of 90 min. Glucose produced at each incubation period was determined for specific activity determination. (Personal communication, Yu, M., 2000.) ,QFXEDWLRQWLPHPLQ 6SHFLILFHQ]\PHDFWLYLW\ &RQWURO P0&D&O LA4154/frame/C04 Page 40 Wednesday, May 17, 2000 3:52 PM © 2001 by CRC Press LLC [...]... liver Studies show that in the liver CCl4 is broken down into reactive free radicals, i.e., ·CCl3 and Cl· (Equation 4. 4) It is suggested that the free radicals, in turn, can damage the liver by reacting with liver cellular components Cyt P450 CCl 4 → ⋅CCl 3 + Cl⋅ © 2001 by CRC Press LLC (4. 4) LA41 54/ frame/C 04 Page 42 Wednesday, May 17, 2000 3:52 PM 42 ENVIRONMENTAL TOXICOLOGY (c) A third example... expression of estrogen-responsiveness genes Steroidal estrogens exert their effects through this change in gene expression (Figure 4. 6) A chemical can alter this receptor-mediated process by a number of mechanisms For example, the chemical can change the level of endogenous estrogen at a particular © 2001 by CRC Press LLC LA41 54/ frame/C 04 Page 44 Wednesday, May 17, 2000 3:52 PM 44 ENVIRONMENTAL TOXICOLOGY... observed in wildlife species have raised concern about potential human health effects © 2001 by CRC Press LLC LA41 54/ frame/C 04 Page 45 Wednesday, May 17, 2000 3:52 PM DAMAGE PROCESS AND ACTION OF TOXICANTS 45 4. 5 REFERENCES AND SUGGESTED READINGS 1 Humble, G.D and Raschke, K., Stomatal opening quantitatively related to potassium transport Evidence from electron probe analysis, Plant Physiol., 48 , 44 7, 1971... exposed to O3 for 4 h, the rats showed increased levels of Cu, Mo, and Zn in the lungs, while these metals were decreased in the liver 4. 4.5 Free Radical-Mediated Reactions A free radical is any molecule with an odd number of electrons Free radicals can occur in both organic and inorganic molecules They are highly reactive, and, therefore, highly unstable and short-lived For example, the half -life (T1/2)... attributable to free radical-induced reactions As they react with the unsaturated bonds of fatty acids and cholesterol, such as those in membranes, free radicals can induce lipid peroxidation This latter process, © 2001 by CRC Press LLC LA41 54/ frame/C 04 Page 43 Wednesday, May 17, 2000 3:52 PM DAMAGE PROCESS AND ACTION OF TOXICANTS 43 + 5+ → 5 5 2 → 52 52 5+ → 522+ 5 Figure 4. 5 Lipid peroxidation... injury, Lab Invest., 47 , 41 2, 1982 14 Fry, D.M and Toone, C.K., DDT-induced feminization of gull embryos, Science, 213, 922, 1981 15 Schultz, T.W et al., Estrogenicity of selected biphenyls evaluated using a recombinant yeast assay, Environ Toxicol Chem., 17, 1727, 1998 16 Hileman, B., Environmental estrogens linked to reproductive abnormalities, cancer, Chem & Eng News, Jan 31, 19 94, 19 4. 6 REVIEW QUESTIONS... because it is a potent inhibitor of aconitase, the enzyme that catalyzes the conversion of citrate into cis-aconitate and then into isocitrate in the Krebs cycle Inhibition of aconitase results in citrate accumulation Impaired cycle functioning occurs, leading to disruption of energy metabolism 4. 4 .4 Secondary Action as a Result of the Presence of a Pollutant The presence of a pollutant in a living system... plants than non-acidified soil? 4 Explain the way in which lead may inhibit an enzyme 5 Explain the way in which fluoride may inhibit an enzyme 6 What is meant by facilitated transport? 7 What does an active transport refer to? What are the characteristics involved in this process? 8 List the three main reactions involved in Phase I reaction © 2001 by CRC Press LLC LA41 54/ frame/C 04 Page 46 Wednesday,...LA41 54/ frame/C 04 Page 41 Wednesday, May 17, 2000 3:52 PM DAMAGE PROCESS AND ACTION OF TOXICANTS 41 (d) A toxic metabolite may depress the activity of an enzyme In this case, enzyme inhibition is not caused by the toxicant per se, but rather by its metabolite... Science, 177, 11 94, 1972 7 Schwimmer, S and Balls, A.K., Isolation and properties of crystalline α-amylase from germinated barley, J Biol Chem., 179, 1063, 1 949 8 Beers, E.P and Duke, S.H., Characterization of α-amylase from shoots and cotyledons of pea (Pisum sativum L.) seedlings, Plant Physiol., 92, 11 54, 1990 9 Sarkar, R.K., Banerjee, A., and Mukherji, S., Effects of toxic concentrations of natrium . with liver cellular components. (4. 4)CCl CCl Cl 43 Cyt P450 → ⋅ + ⋅ LA41 54/ frame/C 04 Page 41 Wednesday, May 17, 2000 3:52 PM © 2001 by CRC Press LLC 42 ENVIRONMENTAL TOXICOLOGY (c) A third. (Equation 4. 3). With the active site blocked, the activity of the enzyme may be depressed or lost. 2 Enz -SH + Pb 2+ → Enz -S- Pb -S- Enz + 2H + (4. 3) It. hydroperoxide. + 5+ → 5 5 2 → 52 52 5+ → 522+5 LA41 54/ frame/C 04 Page 43 Wednesday, May 17, 2000 3:52 PM © 2001 by CRC Press LLC 44 ENVIRONMENTAL TOXICOLOGY site by altering its synthesis,