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Related Experiment Videos

Cholinesterases in rabbit serum.

V Simeon1, E Reiner, M Skrinjarić-Spoljar

  • 1Institute for Medical Research and Occupational Health, University of Zagreb, Croatia, Yugoslavia.

General Pharmacology
|January 1, 1988
PubMed
Summary

Rabbit serum contains three cholinesterases. Two enzymes hydrolyze both acetylthiocholine and butyrylthiocholine, while a third enzyme hydrolyzes only butyrylthiocholine, identified via enzyme kinetics and inhibitor studies.

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Area of Science:

  • Biochemistry
  • Enzymology
  • Pharmacology

Background:

  • Cholinesterases are crucial enzymes in biological systems.
  • Rabbit serum is known to possess cholinesterase activity.
  • Understanding the specific types and properties of these enzymes is important for various applications.

Purpose of the Study:

  • To identify and characterize the cholinesterases present in native rabbit serum.
  • To differentiate the substrate specificities and kinetic properties of these enzymes.
  • To investigate their sensitivity to specific inhibitors and heat inactivation.

Main Methods:

  • Enzyme kinetics analysis using acetylthiocholine and butyrylthiocholine.
  • Heat inactivation kinetics studies.
  • Phosphorylation kinetics with the organophosphate VX.

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  • Enzyme inhibition assays using selective inhibitors (iso-OMPA, eserine, BNPP, BW-284C51).
  • Polyacrylamide gel electrophoresis (PAA-gels) for protein separation.

    • Main Results:

    • Rabbit serum contains three distinct cholinesterase activities.
    • Two enzymes hydrolyzed both acetylthiocholine and butyrylthiocholine, exhibiting properties of acetylcholinesterase (EC 3.1.1.7) and butyrylcholinesterase (EC 3.1.1.8).
    • A third enzyme hydrolyzed only butyrylthiocholine, showing some carboxylesterase (EC 3.1.1.1) characteristics and was least sensitive to VX.

    Conclusions:

    • Rabbit serum possesses a complex cholinesterase profile with multiple enzyme forms.
    • These enzymes can be differentiated by their substrate specificity, kinetic properties, and inhibitor sensitivity.
    • The findings contribute to a better understanding of serum esterase diversity and function.