Carboxylesterase A

The microsomal fraction consists mainly of vesicles (microsomes) derived from the endoplasmic reticulum (smooth and rough). It contains cytochrome P450 and NADPH/cytochrome P450 reductase (collectively the microsomal monooxygenase system), carboxylesterases, A-esterases, epoxide hydrolases, glucuronyl transferases, and other enzymes that metabolize xenobiotics. The 105,000 g supernatant contains soluble enzymes such as glutathione-5-trans-ferases, sulfotransferases, and certain esterases. The 11,000 g supernatant contains all of the types of enzyme listed earlier. 

Fitz-Gerald, C.H., Deeth, H.C., Kitchen, B.J. 1981. The relationship between the levels of free fatty acids, lipoprotein lipase, carboxylesterase, A-acetyl-p-D-glucosaminidase, somatic cell count and other mastitis indices in bovine milk. J. Dairy Res. 48, 253-265. 

Robbi, M. and Beaufay, H. 1992, Topogenesis of carboxylesterases a rat liver isoenzyme ending in -HTEHT-COOH is a secreted protein, Biochem. Biophys. Res. Commun., vol. 183, no. 2, pp. 836-841. 

In an another application, the enantioselectivity of carboxylesterase A (CesA) from Bacillus subtilis towards butyrate and caprilate esters of 1,2-O-isopropylideneglycerol was improved up to 13-fold by structure-guided mutagenesis. The mutagenesis of positions 166 and 182 in CesA yielded novel variants with enhanced enantioselectivity without significant loss of catalytic activity. ... 

Godinho LF, Reis CR, Rozeboom HJ, Dekker FJ, Dijks BW, Poelarends GJ, et al. Enhancement of the enantioselectivity of carboxylesterase A by structure-based mutagenesis.J Biotechnol 2012 158 36-43. 

This information was used to cmistruct a PBPK/PD model in the adult male rat (MirfazaeUan et al. 2006). Godin et al. (2006) examined species differences between rat and human liver microsomal carboxylesterases. A significant species difference was noted in the in vitro biotransformation of deltamethrin, due in part to differences in the rate of hydrolysis by human liver microsomes. Godin et al. (2007) identified the rat and human CYP450 isoforms, and rat serum esterases that metabolize deltamethrin and esfenvalerate. Differences in the rates of hepatic oxidative metabolism were related to expression levels (abundance) of the individual P450 isoforms rather than their specific activity. 

The metabolism of foreign compounds (xenobiotics) often takes place in two consecutive reactions, classically referred to as phases one and two. Phase I is a functionalization of the lipophilic compound that can be used to attach a conjugate in Phase II. The conjugated product is usually sufficiently water-soluble to be excretable into the urine. The most important biotransformations of Phase I are aromatic and aliphatic hydroxylations catalyzed by cytochromes P450. Other Phase I enzymes are for example epoxide hydrolases or carboxylesterases. Typical Phase II enzymes are UDP-glucuronosyltrans-ferases, sulfotransferases, N-acetyltransferases and methyltransferases e.g. thiopurin S-methyltransferase. 

Permethrin, a pyrethrin pesticide, decreased the inhibition of brain cholinesterase activity by methyl parathion, but methyl parathion decreased the LD50 of permethrin when the two pesticides were simultaneously administered to rats (Ortiz et al. 1995). The potentiation of permethrin lethality may be due to the inhibition by methyl parathion of carboxylesterase, which metabolizes permethrin. 

In addition to ester bonds with P (Section 10.2.1, Figures 10.1 and 10.2), some OPs have other ester bonds not involving P, which are readily broken by esteratic hydrolysis to bring about a loss of toxicity. Examples include the two carboxylester bonds of malathion, and the amido bond of dimethoate (Figure 10.2). The two carboxylester bonds of malathion can be cleaved by B-esterase attack, a conversion that provides the basis for the marked selectivity of this compound. Most insects lack an effective carboxylesterase, and for them malathion is highly toxic. Mammals and certain resistant insects, however, possess forms of carboxylesterase that rapidly hydrolyze these bonds, and are accordingly insensitive to malathion toxicity. 

A nnmber of other examples are known in which genetically based resistance was dne to enhanced detoxication of OPs. These include malathion resistance in some stored product pests owing to high carboxylesterase activity, and resistance of strains of the housefly to diazinon due to detoxication by specific forms of a glutathione-S-transferase and monooxygenase (Brooks 1972). 

Satoh, H. et al., Human anti-endoplasmic reticulum antibodies in sera of patients with halothane-induced hepatitis are directed against a trifluoroacetylated carboxylesterase, Proc. Nat. Acad. Sci. USA, 86, 322, 1989. 

In addition, three types of lipophilic conjugates have been found in pyrethroid metabolism studies (Fig. 4). They are cholesterol ester (fenvalerate) [15], glyceride (3-PBacid, a common metabolite of several pyrethroids) [16], and bile acid conjugates (fluvalinate) [17]. It is noteworthy that one isomer out of the four chiral isomers of fenvalerate yields a cholesterol ester conjugate from its acid moiety [15]. This chiral-specific formation of the cholesterol ester has been demonstrated to be mediated by transesterification reactions of carboxylesterase(s) in microsomes, not by any of the three known biosynthetic pathways of endogenous cholesterol esters... 

Munger JS, Shi GP, Mark EA, Chin DT, Gerard C, Chapman HA (1991) A serine esterase released by human alveolar macrophages is closely related to liver microsomal carboxylesterases. J Biol Chem 266 18832-18838... 

Ross MK, Borazjani A, Edwards CC, Potter PM (2006) Hydrolytic metabolism of pyrethroids by human and other mammalian carboxylesterases. Biochem Pharmacol 71 657-669... 

Crow JA, Borazjani A, Potter PM, Ross MK (2007) Hydrolysis of pyrethroids by human and rat tissues examination of intestinal, liver and serum carboxylesterases. Toxicol Appl Pharmacol 221 1-12... 

Urinary proteins were analyzed by SDS-polyacrylamide gel electrophoresis (PAGE), and a 70-kDa protein was identified as the major component of cat urine (Fig. 4.1 A). Comparative analysis of urinary proteins in several other mammals such as humans, mice, dogs, and cattle did not detect a 70-kDa protein. Therefore, the 70-kDa protein was purified from cat urine and characterized by biochemical methods (Miyazaki, Kamiie, Soeta, Taira and Yamashita 2003). Analysis of tissue distribution indicated that the 70-kDa protein is expressed in the kidney in a tissue-specific manner and secreted from the proximal straight tubular cells of the kidney into the urine (Fig. 4.IB). A full-length cDNA for a 70-kDa protein was cloned from a cat kidney cDNA library. The cDNA clone encoded a polypeptide of 545 amino acid residues. The deduced amino acid sequence shared 47% identity with cat carboxylesterase (CES, EC 3.1.1.1), and contained both the CES family protein motif (EDCLY) and a conserved active site motif (GESAG) associated with... 

Cauxin is markedly different from previously reported mammalian CESs in term of urinary excretion. Other mammalian CESs comprise multigene families, and CES isozymes are highly and ubiquitously expressed in tissues such as the brain, liver, kidney, lung, and small intestine (Satoh and Hosokawa 1998). Our work on cauxin was the first description of a carboxylesterase excreted in urine. 

Although proteinuria is often considered to be a pathological event, we demonstrated that this is not the case for the domestic cat. Male cat urine contains a large amount of the mammalian carboxylesterase family member termed cauxin. Cauxin is excreted in a species-, sex-, and age-dependent manner and regulates the production of felinine, a putative pheromone precursor. This finding provides an example of a previously unknown type of proteinuria involved in chemical communication. 

Ecroyd, H., Belghazi, M., Dacheux, J.L., Miyazaki, M., Yamashita, T. and Gatti, J.L. (2006) An epididymal form of cauxin, a carboxylesterase-like enzyme, is present and active in mammalian male reproductive fluids. Biol. Reprod. 74, 439-447. 

Miyazaki, M., Kamiie, K., Soeta, S., Taira, H. and Yamashita, T. (2003) Molecular cloning and characterization of a novel carboxylesterase-like protein that is physiologically present at high concentrations in the mine of domestic cats (Felis catus). Biochem. J. 370, 101-110. 

Miyazaki, M., Yamashita, T., Hosokawa, M., Taira, H. and Suzuki, A. (2006a) Species-, sex-, and age-dependent urinary excretion of cauxin, a mammalian carboxylesterase family. Comp. Biochem. Physiol. B Biochem. Mol. Biol. 145, 270-277. 

Cross-tolerance between disulfoton and another organophosphate, chlorpyrifos, was observed in mice (Costa and Murphy 1983b). Because of this cross-tolerance, a benefit is derived as a result of this interaction. In the same study, propoxur-tolerant mice were tolerant to disulfoton but not vice versa. Propoxur (a carbamate) is metabolized by carboxylesterases, and these enzymes are inhibited in disulfoton-tolerant animals disulfoton-tolerant animals are more susceptible to propoxur and/or carbamate insecticides than are nonpretreated animals. In another study, disulfoton-tolerant rats were tolerant to the cholinergic effects of octamethyl pyrophosphoramide (OMPA) but not parathion (McPhillips 1969a, 1969b). The authors were unable to explain why the insecticides OMPA and parathion caused different effects. 

First generation of topi inhibitors were developed as drugs from camptothecins, a family of compounds derived from wood and bark of the Chinese tree Camptotheca acuminata) [9, 10], Many of these are already in clinical use or clinical trials, including irinotecan, topotecan, exatecan, rubitecan, and lurtotecan. Irinotecan (CPT-11) is bioactivated in liver by carboxylesterase to the active metabolite SN-38, 1000-fold more active [11]. Irinotecan received in 1998 FDA approval for treatment of metastatic colorectal cancer after failure of treatment with 5FU [12],... 

Second, esterases have broad (or even very broad) and overlapping substrate specificities. For example, carboxylesterase (EC 3.1.1.1) also catalyzes reactions characteristic of a number of other hydrolases. The discovery that individual isoenzymes of carboxylesterases may be identical to or closely related to acylglycerol lipase, acylcamitine hydrolase, and palmitoyl-CoA hydrolase (see Sect. 2.4.3) has increased the confusion surrounding esterase classification [59], Many esterases are able to hydrolyze amides, thiolesters,... 

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