Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Recent Progress in <i>N</i>-Acylethanolamine Research: Biological Functions and Metabolism Regulated by Two Distinct <i>N</i>-Acyltransferases: cPLA<sub>2</sub>ε and PLAAT Enzymes.

International journal of molecular sciences·2025
Same author

The PLAAT family as phospholipid-related enzymes.

Progress in lipid research·2025
Same author

PLAAT5 as an N-acyltransferase responsible for the generation of anti-inflammatory N-acylethanolamines in testis.

Biochimica et biophysica acta. Molecular and cell biology of lipids·2024
Same author

PLAAT1 expression triggers fragmentation of mitochondria in an enzyme activity-dependent manner.

Journal of biochemistry·2023
Same author

PLAAT1 deficiency alleviates high-fat diet-induced hepatic lipid accumulation in mice.

FASEB journal : official publication of the Federation of American Societies for Experimental Biology·2023
Same author

GDE7 produces cyclic phosphatidic acid in the ER lumen functioning as a lysophospholipid mediator.

Communications biology·2023

Related Experiment Video

Updated: Aug 27, 2025

Preparation and In Vivo Use of an Activity-based Probe for N-acylethanolamine Acid Amidase
11:01

Preparation and In Vivo Use of an Activity-based Probe for N-acylethanolamine Acid Amidase

Published on: November 23, 2016

9.8K

Assay of NAT Activity.

Toru Uyama1, Natsuo Ueda2

  • 1Department of Biochemistry, Kagawa University School of Medicine, Miki, Kagawa, Japan.

Methods in Molecular Biology (Clifton, N.J.)
|September 24, 2022
PubMed
Summary

This study details the purification of calcium-dependent N-acyltransferase (NAT) and recombinant cPLA2ε and PLAAT-2. Assays were developed to study N-acyltransferase activity in biological systems.

Keywords:
COS-7 cellCa-NATN-AcylphosphatidylethanolamineN-AcyltransferaseNAPEPLAAT familyRadioisotopeRat brainThin-layer chromatographycPLA2ε

More Related Videos

NMR-Based Activity Assays for Determining Compound Inhibition, IC50 Values, Artifactual Activity, and Whole-Cell Activity of Nucleoside Ribohydrolases
10:24

NMR-Based Activity Assays for Determining Compound Inhibition, IC50 Values, Artifactual Activity, and Whole-Cell Activity of Nucleoside Ribohydrolases

Published on: June 30, 2019

10.1K
Analytical Techniques for Assaying Nitric Oxide Bioactivity
11:28

Analytical Techniques for Assaying Nitric Oxide Bioactivity

Published on: June 18, 2012

18.1K

Related Experiment Videos

Last Updated: Aug 27, 2025

Preparation and In Vivo Use of an Activity-based Probe for N-acylethanolamine Acid Amidase
11:01

Preparation and In Vivo Use of an Activity-based Probe for N-acylethanolamine Acid Amidase

Published on: November 23, 2016

9.8K
NMR-Based Activity Assays for Determining Compound Inhibition, IC50 Values, Artifactual Activity, and Whole-Cell Activity of Nucleoside Ribohydrolases
10:24

NMR-Based Activity Assays for Determining Compound Inhibition, IC50 Values, Artifactual Activity, and Whole-Cell Activity of Nucleoside Ribohydrolases

Published on: June 30, 2019

10.1K
Analytical Techniques for Assaying Nitric Oxide Bioactivity
11:28

Analytical Techniques for Assaying Nitric Oxide Bioactivity

Published on: June 18, 2012

18.1K

Area of Science:

  • Biochemistry
  • Enzymology
  • Molecular Biology

Background:

  • N-acyltransferase (NAT) initiates the synthesis of bioactive N-acylethanolamines by transferring acyl chains to phosphatidylethanolamine.
  • Calcium-dependent NAT, identified as cPLA2ε, was previously characterized.
  • Phospholipase A/acyltransferase (PLAAT) family enzymes exhibit calcium-independent NAT activity.

Purpose of the Study:

  • To describe the partial purification of calcium-dependent NAT from rat brain.
  • To detail the purification of recombinant cPLA2ε and PLAAT-2.
  • To outline the methodology for N-acyltransferase assays using radiolabeled substrates.

Main Methods:

  • Partial purification of calcium-dependent NAT from rat brain tissue.
  • Recombinant protein expression and purification of cPLA2ε and PLAAT-2.
  • Development and application of NAT assays utilizing radiolabeled phospholipid substrates.

Main Results:

  • Successful partial purification of Ca2+-dependent NAT from rat brain.
  • Purification of functional recombinant cPLA2ε and PLAAT-2 enzymes.
  • Established protocols for NAT activity assays.

Conclusions:

  • The study provides methods for purifying and assaying both calcium-dependent and calcium-independent N-acyltransferases.
  • These methods facilitate further research into the biosynthesis of N-acylethanolamines.
  • Characterization of cPLA2ε and PLAAT family enzymes is crucial for understanding lipid signaling pathways.