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

Aerobic catabolism of isobutylene by <i>Mycolicibacterium</i> sp. ELW1 requires inducible, plasmid-borne <i>ibc</i> genes.

Applied and environmental microbiology·2026
Same author

Characterization of an isobutylene epoxide hydrolase (IbcK) from the isobutylene-catabolizing bacterium <i>Mycolicibacterium</i> sp. ELW1.

Applied and environmental microbiology·2025
Same author

Diyne inactivators and activity-based fluorescent labeling of phenol hydroxylase in Pseudomonas sp. CF600.

FEMS microbiology letters·2023
Same author

Single-well push-pull tests evaluating isobutane as a primary substrate for promoting in situ cometabolic biotransformation reactions.

Biodegradation·2022
Same author

Draft Genome Sequences of Four Aerobic Isobutane-Metabolizing Bacteria.

Microbiology resource announcements·2021
Same author

Co-encapsulation of slow release compounds and Rhodococcus rhodochrous ATCC 21198 in gellan gum beads to promote the long-term aerobic cometabolic transformation of 1,1,1-trichloroethane, cis-1,2-dichloroethene and 1,4-dioxane.

Environmental science. Processes & impacts·2020

Related Experiment Video

Updated: Jun 16, 2025

Anaerobic Protein Purification and Kinetic Analysis via Oxygen Electrode for Studying DesB Dioxygenase Activity and Inhibition
08:31

Anaerobic Protein Purification and Kinetic Analysis via Oxygen Electrode for Studying DesB Dioxygenase Activity and Inhibition

Published on: October 3, 2018

8.5K

Activity-Based Protein Profiling of Bacterial Monooxygenases.

John B Joyce1, Michael R Hyman2

  • 1Department of Plant and Microbial Biology, North Carolina State University, Raleigh, NC, USA.

Methods in Molecular Biology (Clifton, N.J.)
|June 14, 2025
PubMed
Summary
This summary is machine-generated.

Activity-based protein profiling (ABPP) offers a new way to study bacterial monooxygenases, including ammonia monooxygenase (AMO). This method helps characterize these key enzymes, even when purification is difficult.

Keywords:
Activity-based protein profilingDiyne probeMonooxygenasesNear-infrared (NIR) fluorescent imagingSDS-PAGE

More Related Videos

Benchtop Immobilized Metal Affinity Chromatography, Reconstitution and Assay of a Polyhistidine Tagged Metalloenzyme for the Undergraduate Laboratory
08:02

Benchtop Immobilized Metal Affinity Chromatography, Reconstitution and Assay of a Polyhistidine Tagged Metalloenzyme for the Undergraduate Laboratory

Published on: August 23, 2018

17.4K
Monitoring the Reductive and Oxidative Half-Reactions of a Flavin-Dependent Monooxygenase using Stopped-Flow Spectrophotometry
12:08

Monitoring the Reductive and Oxidative Half-Reactions of a Flavin-Dependent Monooxygenase using Stopped-Flow Spectrophotometry

Published on: March 18, 2012

15.1K

Related Experiment Videos

Last Updated: Jun 16, 2025

Anaerobic Protein Purification and Kinetic Analysis via Oxygen Electrode for Studying DesB Dioxygenase Activity and Inhibition
08:31

Anaerobic Protein Purification and Kinetic Analysis via Oxygen Electrode for Studying DesB Dioxygenase Activity and Inhibition

Published on: October 3, 2018

8.5K
Benchtop Immobilized Metal Affinity Chromatography, Reconstitution and Assay of a Polyhistidine Tagged Metalloenzyme for the Undergraduate Laboratory
08:02

Benchtop Immobilized Metal Affinity Chromatography, Reconstitution and Assay of a Polyhistidine Tagged Metalloenzyme for the Undergraduate Laboratory

Published on: August 23, 2018

17.4K
Monitoring the Reductive and Oxidative Half-Reactions of a Flavin-Dependent Monooxygenase using Stopped-Flow Spectrophotometry
12:08

Monitoring the Reductive and Oxidative Half-Reactions of a Flavin-Dependent Monooxygenase using Stopped-Flow Spectrophotometry

Published on: March 18, 2012

15.1K

Area of Science:

  • Biochemistry
  • Proteomics
  • Environmental Microbiology

Background:

  • Bacterial monooxygenases are crucial for biogeochemical cycles.
  • Some monooxygenases, like ammonia monooxygenase (AMO), are challenging to characterize using traditional purification methods.
  • Activity-based protein profiling (ABPP) provides a powerful alternative for studying enzyme activity.

Purpose of the Study:

  • To present a novel 2-step ABPP method for characterizing bacterial monooxygenases.
  • To focus on SDS-PAGE-based detection of fluorescently labeled monooxygenase components.
  • To highlight factors influencing ABPP for monooxygenase quantification and cellular localization.

Main Methods:

  • Developed and applied a 2-step activity-based protein profiling (ABPP) strategy.
  • Utilized SDS-PAGE for the detection of fluorescently labeled monooxygenase proteins.
  • Investigated features impacting quantification and cellular detection of target enzymes.

Main Results:

  • Successfully characterized ammonia monooxygenase (AMO) and other bacterial monooxygenases using the developed ABPP method.
  • Demonstrated the utility of SDS-PAGE for visualizing activity-based labeled enzyme components.
  • Identified key analytical considerations for quantitative ABPP of monooxygenases.

Conclusions:

  • The described 2-step ABPP method is effective for characterizing bacterial monooxygenases, including AMO.
  • SDS-PAGE-based detection is a viable approach for analyzing activity-labeled enzymes.
  • This ABPP approach enhances the study of enzymes vital to biogeochemical processes.