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A Methodology for Detection and Quantification of Esterase Activity

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Capillary Electrophoresis of Biomolecules

Part of the book series: Methods in Molecular Biology ((MIMB,volume 984))

Abstract

Carboxylesterases are important enzymes for xenobiotic metabolism and are receiving increasing attention in the context of cancer therapies. Quantification of individual carboxylesterase activity is important since protein levels do not always correlate to activity and significant interorgan, interindividual, and interspecies variations exist. Here we present a methodology enabling the specific quantification of carboxylesterase 2 activity in a pool of other esterases. Method applicability is illustrated for the evaluation of interspecies variation and for activity assessment of transfected cell extracts. The methodology can easily be adapted to the evaluation of other esterases upon careful selection of adequate substrates and/or specific inhibitors.

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References

  1. Imai T, Ohura K (2010) The role of intestinal carboxylesterase in the oral absorption of prodrugs. Curr Drug Metab 11:793–805

    Article  PubMed  CAS  Google Scholar 

  2. Satoh T, Hosokawa M (2006) Structure, function and regulation of carboxylesterases. Chem Biol Interact 162:195–211

    Article  PubMed  CAS  Google Scholar 

  3. Crow JA, Borazjani A, Potter PM et al (2007) Hydrolysis of pyrethroids by human and rat tissues: examination of intestinal, liver and serum carboxylesterases. Toxicol Appl Pharmacol 221:1–12

    Article  PubMed  CAS  Google Scholar 

  4. Kubo T, Kim SR, Sai K et al (2005) Functional characterization of three naturally occurring single nucleotide polymorphisms in the CES2 gene encoding carboxylesterase 2 (HCE-2). Drug Metab Dispos 33:1482–1487

    Article  PubMed  CAS  Google Scholar 

  5. Lamego J, Coroadinha AS, Simplício AL (2011) Detection and quantification of carboxylesterase 2 activity by capillary electrophoresis. Anal Chem 83:881–887

    Article  PubMed  CAS  Google Scholar 

  6. Tang ZM, Wang ZY, Kang JW (2007) Screening of acetylcholinesterase inhibitors in natural extracts by CE with electrophoretically m­ediated microanalysis technique. Electrophoresis 28:360–365

    Article  PubMed  CAS  Google Scholar 

  7. Pindel EV, Kedishvili NY, Abraham TL et al (1997) Purification and cloning of a broad substrate specificity human liver carboxylesterase that catalyzes the hydrolysis of cocaine and heroin. J Biol Chem 272:14769–14775

    Article  PubMed  CAS  Google Scholar 

  8. Li B, Sedlacek M, Manoharan I et al (2005) Butyrylcholinesterase, paraoxonase, and albumin esterase, but not carboxylesterase, are present in human plasma. Biochem Pharmacol 70:1673–1684

    Article  PubMed  CAS  Google Scholar 

  9. Christensen JM, Stalker D (1991) Ibuprofen piconol hydrolysis in vitro in plasma, whole blood, and serum using different anticoagulants. J Pharm Sci 80:29–31

    Article  PubMed  CAS  Google Scholar 

  10. Ross MK, Borazjani A (2007) Enzymatic Activity of Human Carboxylesterases. Curr Protoc Toxicol 33:4.24.1–4.24.14

    Google Scholar 

  11. Williams ET, Ehsani ME, Wang X et al (2008) Effect of buffer components and carrier solvents on in vitro activity of recombinant human carboxylesterases. J Pharmacol Toxicol Methods 57:138–144

    Article  PubMed  CAS  Google Scholar 

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Acknowledgment

This work was funded by Fundação para a Ciência e Tecnologia, Portugal (SFRH/BD/44025/2008, PTDC/EBB-BIO/111530/2009, PEst-OE/EQB/LA0004/2011).

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Correspondence to Ana L. Simplício .

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Simplício, A.L., Coroadinha, A.S., Gilmer, J.F., Lamego, J. (2013). A Methodology for Detection and Quantification of Esterase Activity. In: Volpi, N., Maccari, F. (eds) Capillary Electrophoresis of Biomolecules. Methods in Molecular Biology, vol 984. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-296-4_22

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  • DOI: https://doi.org/10.1007/978-1-62703-296-4_22

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-295-7

  • Online ISBN: 978-1-62703-296-4

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