Relationship between sequence conservation and three‐dimensional structure in a large family of esterases, lipases, and related proteins

Cygler, Mirosław; Schrag, Joseph D.; Sussman, Joel L.; Harel, Michal; Silman, Israel; Gentry, Mary K.; Doctor, Bhupendra P.

doi:10.1002/pro.5560020309

Cited by 549 publications

(372 citation statements)

References 66 publications

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“…Although its functional domains remain uncharacterized, ASPA has been referred to as a member of the α/β-hydrolase superfamily of enzymes because of its enzymatic inhibition by diisopropyl fluorophosphates, which suggests a catalytic serine, and because of its local sequence homology to catalytic cores in esterases (Cygler et al 1993,Matalon & Michals-Matalon 1999. However, ASPA lacks an invariant serine, a candidate for a catalytic residue, and an alignment study revealed minimal homology of ASPA with serine proteases (Makarova & Grishin 1999,Cygler et al 1993).…”

Section: Introductionmentioning

confidence: 99%

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Mutational analysis of aspartoacylase: Implications for Canavan Disease

et al. 2007

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Mutations that result in undetectable activity of aspartoacylase, which catalyzes the deacetylation of N-acetyl-L-aspartate, correlate with Canavan Disease, a neurodegenerative disorder usually fatal during childhood. The underlying biochemical mechanisms of how these mutations ablate activity are poorly understood. Therefore, we developed and tested a three-dimensional homology model of aspartoacylase based on zinc dependent carboxypeptidase A. Mutations of the putative zinc-binding residues (H21G, E24D/G, and H116G), the general proton donor (E178A), and mutants designed to switch the order of the zinc-binding residues (H21E/E24H and E24H/H116E) yielded wild-type aspartoacylase protein levels and undetectable ASPA activity. Mutations that affect substrate carboxyl binding (R71N) and transition state stabilization (R63N) also yielded wild-type aspartoacylase protein levels and undetectable aspartoacylase activity. Alanine substitutions of Cys124 and Cys152, residues indicated by homology modeling to be in close proximity and in the proper orientation for disulfide bonding, yielded reduced ASPA protein and activity levels. Finally, expression of several previously tested (E24G, D68A, C152W, E214X, D249V, E285A, and A305E) and untested (H21P, A57T, I143T, P183H, M195R, K213E/G274R, G274R, and F295S) Canavan Disease mutations resulted in undetectable enzyme activity, and only E285A and P183H showed wild-type aspartoacylase protein levels. These results show that aspartoacylase is a member of the caboxypeptidase A family and offer novel explanations for most loss-of-function aspartoacylase mutations associated with Canavan Disease.

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Section: Introductionmentioning

confidence: 99%

“…However, ASPA lacks an invariant serine, a candidate for a catalytic residue, and an alignment study revealed minimal homology of ASPA with serine proteases (Makarova & Grishin 1999,Cygler et al 1993). …”

Section: Introductionmentioning

confidence: 99%

Mutational analysis of aspartoacylase: Implications for Canavan Disease

et al. 2007

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“…38 TcAChE is homologous to several lipases and esterases, resulting in similar threedimensional structures for all of them. 39 However, this folding pattern (a β-sheet surrounded by α-helices) is also found in hydrolases that bear no similarity to TcAChE or among themselves. This fold, first identified in the 1990's, is designated α/β hydrolase.…”

Section: Ache Structurementioning

confidence: 98%

“…40 It is believed that, in TcAChE, this folding is kept by four salt bridges. 39 Indepth descriptions of structural details and disulfide-linkage patterns, as well as comparisons with structures of other enzymes, can be found elsewhere. 24,28,38,39 …”

Section: Ache Structurementioning

confidence: 99%

Organophosphorus compounds as chemical warfare agents: a review

Delfino¹,

Ribeiro²,

Figueroa‐Villar³

2009

J. Braz. Chem. Soc.

231

178

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Os agentes de guerra química constituem uma das maiores ameaças do mundo moderno. Dentre estes, destacam-se os agentes neurotóxicos, em virtude de sua alta letalidade e periculosidade. Eles são compostos organofosforados que atuam pela inibição da enzima acetilcolinesterase, a qual é fundamental no processo de transmissão de impulsos nervosos. Existem várias formas de tratamento para a intoxicação por organofosforados, mas nenhuma delas é eficaz contra todos os agentes conhecidos ou contra todos os seus efeitos. Esta revisão tem como foco o uso de compostos organofosforados como agentes neurotóxicos de guerra química. Após uma breve introdução histórica, será feita uma discussão sobre as principais características estruturais e biológicas da acetilcolinesterase, seguida por uma revisão das propriedades dos compostos organofosforados e da sua aplicação como agentes de guerra química. Por fim, serão discutidas as formas de tratamento contra estes agentes, com ênfase nas oximas usadas para reativar a acetilcolinesterase inibida.Chemical warfare agents constitute one of the greatest threats in the modern world. Among them, the neurotoxic agents are of special interest due to their high lethality and danger. Neurotoxic agents are organophosphorus compounds that act by inhibiting the enzyme acetylcholinesterase, which is fundamental for the control of transmission of nervous impulses. There are several ways of treating intoxication by organophosphorus compounds, but none of them is efficient against all the known neurotoxic agents or against all of their effects. This review focus on the use of organophosphorus compounds as neurotoxic chemical warfare agents. After a brief historical introduction, it will be done a discussion about the structural and biological characteristics of acetylcholinesterase, followed by a review of the properties of organophosphorus compounds and their application as chemical warfare agents. Finally, the ways of treatment against intoxication with these agents will be discussed, with emphasis on the oximes used for reactivating the inhibited acetylcholinesterase.

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“…[1][2][3] They are responsible for the hydrolysis of numerous exogenous and endogenous ester-containing compounds 4,5 and have become of increased interest due to their utility in the activation of prodrugs and metabolism of softdrugs, 6 including the chemotherapeutic agent CPT-11 7 and the anti-influenza viral agent oseltamivir 8 . CaEs are also important in agrochemical research because they detoxify pyrethroid, organophosphate and carbamate insecticides.…”

Section: Introductionmentioning

confidence: 99%

Influence of sulfur oxidation state and steric bulk upon trifluoromethyl ketone (TFK) binding kinetics to carboxylesterases and fatty acid amide hydrolase (FAAH)

Wheelock

Nishi

Ying

et al. 2008

Bioorganic & Medicinal Chemistry

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Carboxylesterases metabolize numerous exogenous and endogenous ester-containing compounds including the chemotherapeutic agent CPT-11, anti-influenza viral agent oseltamivir and many agrochemicals. Trifluoromethyl ketone (TFK)-containing compounds with a sulfur atom beta to the ketone moiety are some of the most potent carboxylesterase and amidase inhibitors identified to date. This study examined the effects of alkyl chain length (i.e., steric effects) and sulfur oxidation state upon TFK inhibitor potency (IC 50 ) and binding kinetics (k i ). The selective carboxylesterase inhibitor benzil was used as a non-TFK containing control. These effects were examined using two commercial esterases (porcine and rabbit liver esterase) and two human recombinant esterases (hCE-1 and hCE-2) as well as human recombinant fatty acid amide hydrolase (FAAH). In addition, the inhibition mechanism was examined using a combination of 1 H NMR, X-ray crystallography and ab initio calculations. Overall, the data show that while sulfur oxidation state profoundly affects both inhibitor potency and binding kinetics, the steric effects dominate and override the contributions of sulfur oxidation. In addition, the data suggest that inclusion of a sulfur atom beta to the ketone contributes an increase (~5-fold) in inhibitor potency due to effects upon ketone hydration and/or intramolecular hydrogen bond formation. These results provide further information on the nature of the TFK binding interaction and will be useful in increasing our understanding of this basic biochemical process.

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Relationship between sequence conservation and three‐dimensional structure in a large family of esterases, lipases, and related proteins

Cited by 549 publications

References 66 publications

Mutational analysis of aspartoacylase: Implications for Canavan Disease

Mutational analysis of aspartoacylase: Implications for Canavan Disease

Organophosphorus compounds as chemical warfare agents: a review

Influence of sulfur oxidation state and steric bulk upon trifluoromethyl ketone (TFK) binding kinetics to carboxylesterases and fatty acid amide hydrolase (FAAH)

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