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

Phosphorylation01:02

Phosphorylation

The addition or removal of phosphate groups from proteins is the most common chemical modification that regulates cellular processes. These modifications can affect the structure, activity, stability, and localization of proteins within cells as well as their interactions with other proteins.
During phosphorylation, protein kinases transfer the terminal phosphate group of ATP to specific amino acid side chains of substrate proteins. Serine, threonine, and tyrosine are the most commonly...
Phosphorylation01:02

Phosphorylation

The addition or removal of phosphate groups from proteins is the most common chemical modification that regulates cellular processes. These modifications can affect the structure, activity, stability, and localization of proteins within cells as well as their interactions with other proteins.
During phosphorylation, protein kinases transfer the terminal phosphate group of ATP to specific amino acid side chains of substrate proteins. Serine, threonine, and tyrosine are the most commonly...
Calmodulin-dependent Signaling01:16

Calmodulin-dependent Signaling

Calmodulin (CaM) is a calcium-binding protein in eukaryotes that controls various calcium-regulated cellular processes. It has four calcium-binding sites that bind calcium to form the calcium-calmodulin ( Ca2+-CaM) complex. GPCR stimulation increases the calcium levels in the cells that bind to CaM and induces a conformational change.
The Ca2+-CaM complex does not have enzymatic activity by itself. Instead, the complex binds downstream target proteins, including membrane proteins or enzymes,...
Protein Kinases and Phosphatases02:54

Protein Kinases and Phosphatases

Proteins undergo chemical modifications that trigger changes in the charge, structure, and conformation of the proteins. Phosphorylation, acetylation, glycosylation, nitrosylation, ubiquitination, lipidation, methylation, and proteolysis are various protein modifications that regulate protein activity. Such modifications are usually enzyme-driven.
Protein kinases
Many proteins in the cell are regulated by phosphorylation, the addition of a phosphate group. A family of enzymes called kinases...
Protein Kinases and Phosphatases02:54

Protein Kinases and Phosphatases

Proteins undergo chemical modifications that trigger changes in the charge, structure, and conformation of the proteins. Phosphorylation, acetylation, glycosylation, nitrosylation, ubiquitination, lipidation, methylation, and proteolysis are various protein modifications that regulate protein activity. Such modifications are usually enzyme-driven.
Protein kinases
Many proteins in the cell are regulated by phosphorylation, the addition of a phosphate group. A family of enzymes called kinases...
Amplifying Signals via Enzymatic Cascade01:22

Amplifying Signals via Enzymatic Cascade

When a ligand binds to a cell-surface receptor, the receptor's intracellular domain changes shape, which may either activate its enzyme function or allow its binding to other molecules. The initial signal is amplified by most signal transduction pathways. This means that a single ligand molecule can activate multiple molecules of a downstream target. Proteins that relay a signal are most commonly phosphorylated at one or more sites, activating or inactivating the protein. Kinases catalyze the...

You might also read

Related Articles

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

Sort by
Same author

How hyperglycemia promotes ocular diseases: An overview of molecular mechanisms.

Experimental eye research·2026
Same author

Role of the Wnt/β-Catenin Signaling Pathway in Mediating Outer Root Sheath Stem Cells to Promote Hair Follicle Regeneration and Skin Wound Healing in Mice.

Cells·2026
Same author

Variable selection in causal semiparametric transformation models with all-or-nothing treatment compliance.

Lifetime data analysis·2026
Same author

Oridonin Suppresses Colorectal Cancer Growth In Vitro and In Vivo: Evidence from Integrated Transcriptomic and Proteomic Profiling.

Current issues in molecular biology·2026
Same author

Interpretable Deep Regression Models With Interval-Censored Failure Time Data.

Statistics in medicine·2026
Same author

Integrative machine learning reveals interplay between epithelial and endothelial cells elevates mixed heavy metal-related colorectal cancer risk.

Journal of biomedical research·2026
Same journal

Vancomycin-Mediated Binding of DNA Origami Nanostructures to Gram-Positive and Gram-Negative Bacteria.

Chembiochem : a European journal of chemical biology·2026
Same journal

Mutasynthesis and Antibiotic Activity of Mupirocin Analogues.

Chembiochem : a European journal of chemical biology·2026
Same journal

Pressure-Dependent Aromatic Ring Flips Reveal Variable Transition-State Volume and Compressibility Among Structural Regions of BPTI.

Chembiochem : a European journal of chemical biology·2026
Same journal

Eyes Toward the Clinic: Selective Inhibition and Degradation Approaches to Bromodomain-Containing Proteins.

Chembiochem : a European journal of chemical biology·2026
Same journal

Contrast Agents for Enhanced Bioimaging: A Comprehensive Review.

Chembiochem : a European journal of chemical biology·2026
Same journal

RNAGEN: A Generative Adversarial Network-Based Model to Generate Synthetic RNA Sequences to Target Proteins.

Chembiochem : a European journal of chemical biology·2026
See all related articles

Related Experiment Video

Updated: May 28, 2026

An Optimized Single-Molecule Pull-Down Assay for Quantification of Protein Phosphorylation
07:45

An Optimized Single-Molecule Pull-Down Assay for Quantification of Protein Phosphorylation

Published on: June 6, 2022

Smallest genetically encoded phosphorylation status sensor.

Shuwei Li1

  • 1Institute for Bioscience and Biotechnology Research and Department of Chemistry and Biochemistry, University of Maryland College Park, 9600 Gudelsky Drive, Rockville, MD 20850, USA. sli@umd.edu

Chembiochem : a European Journal of Chemical Biology
|October 18, 2011
PubMed
Summary
This summary is machine-generated.

Researchers developed a novel, environment-sensitive fluorescent amino acid, L-(7-hydroxycoumarin-4-yl) ethylglycine (7HC). This smallest phosphorylation sensor can be genetically encoded, enabling sensitive detection of phosphorylation events with minimal protein disruption.

More Related Videos

Oligopeptide Competition Assay for Phosphorylation Site Determination
09:16

Oligopeptide Competition Assay for Phosphorylation Site Determination

Published on: May 18, 2017

Related Experiment Videos

Last Updated: May 28, 2026

An Optimized Single-Molecule Pull-Down Assay for Quantification of Protein Phosphorylation
07:45

An Optimized Single-Molecule Pull-Down Assay for Quantification of Protein Phosphorylation

Published on: June 6, 2022

Oligopeptide Competition Assay for Phosphorylation Site Determination
09:16

Oligopeptide Competition Assay for Phosphorylation Site Determination

Published on: May 18, 2017

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Biotechnology

Background:

  • Phosphorylation is a critical post-translational modification regulating numerous cellular processes.
  • Existing phosphorylation sensors can be large, potentially interfering with protein structure and function.
  • There is a need for small, minimally invasive sensors for studying phosphorylation dynamics.

Purpose of the Study:

  • To develop and characterize a novel, genetically encodable, environment-sensitive fluorescent amino acid for phosphorylation detection.
  • To evaluate the utility of this amino acid as a small, minimally disruptive phosphorylation sensor.

Main Methods:

  • Genetic encoding of L-(7-hydroxycoumarin-4-yl) ethylglycine (7HC) in E. coli using a suppressor tRNA and cognate synthetase.
  • Incorporation of 7HC into full-length proteins.
  • Demonstration of 7HC's fluorescence changes upon nearby phosphorylation events.

Main Results:

  • L-(7-hydroxycoumarin-4-yl) ethylglycine (7HC) was successfully genetically encoded in E. coli.
  • 7HC was incorporated into full-length proteins without significant structural or functional perturbation.
  • The incorporated 7HC demonstrated environment-sensitive fluorescence, enabling detection of phosphorylation events.

Conclusions:

  • 7HC serves as the smallest genetically encodable phosphorylation sensor to date.
  • This sensor allows for sensitive detection of phosphorylation with minimal impact on protein biology.
  • 7HC offers a promising new tool for studying phosphorylation in various biological contexts.