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 Experiment Video

Updated: Jul 13, 2026

A Fluorescence-based Assay of Phospholipid Scramblase Activity
09:52

A Fluorescence-based Assay of Phospholipid Scramblase Activity

Published on: September 20, 2016

Probing phospholipase a(2) with fluorescent phospholipid substrates.

Oliver Wichmann1, Michael H Gelb, Carsten Schultz

  • 1European Molecular Biology Laboratorium, Gene Expression Programme, Meyerhofstrasse 1, 69117 Heidelberg, Germany.

Chembiochem : a European Journal of Chemical Biology
|July 31, 2007
PubMed
Summary

Researchers modified a sensor (PENN) to detect specific phospholipase A(2) (PLA(2)) activities. Modified sensors with double bonds showed altered performance, highlighting the importance of bioactivation for cellular detection.

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

Identifying Novel Estrogenic Mitochondrial Targets in Hypothalamic Proopiomelanocortin Neurons by Chemoproteomics.

bioRxiv : the preprint server for biology·2026
Same author

Decoding protein-phospholipid interaction networks in cancer: the role of acyl-chain remodeling.

RSC chemical biology·2026
Same author

High-resolution phosphoinositide analysis.

Methods in enzymology·2026
Same author

An Integrated Method for Profiling Lipid-Protein Interactions Using Multifunctional Lipid Probes.

bioRxiv : the preprint server for biology·2026
Same author

Emerging roles of lipids in the flavivirus life cycle.

Trends in microbiology·2026
Same author

SputOMICs identifies common and distinct markers in cystic fibrosis and chronic obstructive pulmonary disease.

Scientific reports·2025

Area of Science:

  • Biochemistry and Molecular Biology
  • Chemical Biology
  • Cellular Imaging and Sensing

Background:

  • The Foerster resonance energy transfer (FRET)-based sensor PENN is utilized for measuring intracellular phospholipase A(2) (PLA(2)) activity in live cells and organisms.
  • Modifying the sn-2 fatty acid of the sensor is a strategy to target specific PLA(2) isoforms.
  • Arachidonic acid serves as a reference structure for designing modified fatty acids.

Purpose of the Study:

  • To develop modified PENN sensors capable of detecting particular phospholipase A(2) isoforms.
  • To investigate the impact of altering the sn-2 fatty acid, specifically the number of double bonds, on sensor activity and specificity.
  • To evaluate the necessity of bioactivation for sensor performance in living cells.

Main Methods:

More Related Videos

Fluorescence-Based Measurements of Phosphatidylserine/Phosphatidylinositol 4-Phosphate Exchange Between Membranes
08:49

Fluorescence-Based Measurements of Phosphatidylserine/Phosphatidylinositol 4-Phosphate Exchange Between Membranes

Published on: March 14, 2021

Defining Substrate Specificities for Lipase and Phospholipase Candidates
08:59

Defining Substrate Specificities for Lipase and Phospholipase Candidates

Published on: November 23, 2016

Related Experiment Videos

Last Updated: Jul 13, 2026

A Fluorescence-based Assay of Phospholipid Scramblase Activity
09:52

A Fluorescence-based Assay of Phospholipid Scramblase Activity

Published on: September 20, 2016

Fluorescence-Based Measurements of Phosphatidylserine/Phosphatidylinositol 4-Phosphate Exchange Between Membranes
08:49

Fluorescence-Based Measurements of Phosphatidylserine/Phosphatidylinositol 4-Phosphate Exchange Between Membranes

Published on: March 14, 2021

Defining Substrate Specificities for Lipase and Phospholipase Candidates
08:59

Defining Substrate Specificities for Lipase and Phospholipase Candidates

Published on: November 23, 2016

  • Synthesis of arachidonic acid-mimicking fatty acids with one or three Z double bonds using copper-mediated coupling reactions.
  • Preparation of novel unsaturated, doubly labeled phosphatidylethanolamine derivatives (PENN2, PENN3) and a saturated derivative (PENN).
  • In vitro enzymatic assays using various secretory phospholipase A(2) (sPLA(2)) isoforms (Group V sPLA(2), hog pancreas sPLA(2)).
  • Cell-based assays in HeLa cells using S-acetylthioethyl (SATE)-activated and membrane-permeant sensor versions.

Main Results:

  • Probes with a single double bond at the 5-position demonstrated favorable substrate properties for secretory PLA(2)s.
  • The sensor PENN2 (one double bond) was preferentially cleaved by Group V sPLA(2), while its O-methyl derivative PMNN2 was best accepted by hog pancreas sPLA(2).
  • Bioactivation using S-acetylthioethyl (SATE) groups was confirmed as essential for probe performance in living cells.
  • Surprisingly, membrane-permeant PENN2 and PENN3 sensors with double bonds showed minimal cleavage in HeLa cells, whereas the saturated PENN sensor was readily cleaved.

Conclusions:

  • Modification of the sn-2 fatty acid in PENN sensors can influence their substrate specificity towards different PLA(2) isoforms.
  • The presence and position of double bonds in the fatty acid chain significantly impact sensor recognition and cleavage by specific PLA(2)s.
  • Bioactivation is critical for the effective cellular uptake and activity of these FRET-based sensors, and structural modifications can unexpectedly reduce cellular performance.