Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

Enzymes

Enzymes

Inside living organisms, enzymes act as catalysts for many biochemical reactions involved in cellular metabolism. The role of enzymes is to reduce the activation energies of biochemical reactions by forming complexes with its substrates. The lowering of activation energies favor an increase in the rates of biochemical reactions.
Enzyme deficiencies can often translate into life-threatening diseases. For example, a genetic abnormality resulting in the deficiency of the enzyme G6PD...

Biosynthesis in Bacteria

Biosynthesis in Bacteria

Biosynthesis in bacteria is a fundamental anabolic process that generates essential macromolecules, including proteins, nucleic acids, lipids, and polysaccharides. These macromolecules are critical for cellular growth, replication, and function. The process is tightly regulated and energetically linked to catabolic pathways to ensure optimal resource utilization.Biosynthetic pathways begin with precursor metabolites such as pyruvate, acetyl-CoA, and glucose-6-phosphate derived from glycolysis,...

Biosynthesis of Polysaccharides

Biosynthesis of Polysaccharides

Polysaccharides such as glycogen and starch are synthesized from nucleoside diphosphate sugars, primarily uridine diphosphate glucose (UDPG) and adenosine diphosphate glucose (ADPG). These activated glucose donors act as key intermediates in carbohydrate metabolism and biosynthesis. UDPG primarily involves glycogen synthesis in animals and many bacteria, while ADPG plays a fundamental role in starch synthesis in plants and certain bacteria.UDPG is formed when glucose-1-phosphate reacts with...

Biosynthesis of Lipids

Biosynthesis of Lipids

Microbial membranes exhibit remarkable diversity in lipid composition, reflecting evolutionary adaptations to various environmental conditions. The three domains of life—Bacteria, Archaea, and Eukarya—synthesize membrane lipids through distinct biosynthetic pathways, leading to fundamental structural differences that impact membrane stability, function, and adaptability.Fatty Acid-Based Lipids in Bacteria and EukaryaBacteria and eukaryotes share a common fatty acid biosynthesis...

Biosynthesis of Nucleic Acids

Biosynthesis of Nucleic Acids

Nucleic acid biosynthesis is a fundamental biochemical process that produces the purine and pyrimidine nucleotides essential for DNA and RNA synthesis. This pathway maintains a balanced nucleotide pool, preventing imbalances that could jeopardize genetic integrity and cellular function. Given the crucial role of nucleotides, their synthesis is tightly regulated to ensure proper cellular homeostasis.Purine BiosynthesisThe biosynthesis of purine nucleotides begins with ribose-5-phosphate, a...

Enzyme Kinetics

Enzyme Kinetics

Enzymes speed up reactions by lowering the activation energy of the reactants. The speed at which the enzyme turns reactants into products is called the rate of reaction. Several factors impact the rate of reaction, including the number of available reactants. Enzyme kinetics is the study of how an enzyme changes the rate of a reaction.
Scientists typically study enzyme kinetics with a fixed amount of enzyme in the controlled environment of a test tube. When more reactant, or substrate, is...

You might also read

Related Articles

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

Sort by

Same author

Development of GS-441524 Derivatives as Potent SARS-CoV-2 Mac1 Inhibitors via a Direct-to-Biology Approach.

bioRxiv : the preprint server for biology·2026

Same author

A ratiometric fluorescent reporter of mitochondrial sodium.

Nature chemical biology·2026

Same author

SIRT2 mediates integrated stress response by deacetylating and stabilizing 4E-BP1 to suppress translation.

EMBO reports·2026

Same author

Coenzyme A is a redox sensing cofactor for malic enzyme 2 regulating oxidative stress and mitochondrial metabolism.

bioRxiv : the preprint server for biology·2026

Same author

Author Correction: Sirtuin 2 inhibits global protein synthesis via Rheb-GTPase degradation.

EMBO reports·2026

Same author

A High-throughput Fluorescence Polarization Assay for Screening Sirtuin Inhibitors.

bioRxiv : the preprint server for biology·2026

Search research articles

Related Experiment Video

Updated: Feb 6, 2026

Free Radicals in Chemical Biology: from Chemical Behavior to Biomarker Development

Free Radicals in Chemical Biology: from Chemical Behavior to Biomarker Development

Published on: April 15, 2013

Methods for Studying the Radical SAM Enzymes in Diphthamide Biosynthesis.

Min Dong¹, Yugang Zhang¹, Hening Lin²

¹Department of Chemistry and Chemical Biology, Cornell University, Ithaca, NY, United States.

Methods in Enzymology

|August 12, 2018

Summary

This study investigates unique radical S-adenosylmethionine (SAM) enzymes involved in diphthamide biosynthesis. Researchers developed methods to prepare, characterize, and assay these enzymes critical for modifying elongation factor 2 (EF2).

Keywords:

Diphthamide Fe–S cluster Radical SAM enzymes

More Related Videos

New Methods to Study Gustatory Coding

New Methods to Study Gustatory Coding

Published on: June 29, 2017

Co-immunoprecipitation Assay for Studying Functional Interactions Between Receptors and Enzymes

Co-immunoprecipitation Assay for Studying Functional Interactions Between Receptors and Enzymes

Published on: September 28, 2018

Related Experiment Videos

Last Updated: Feb 6, 2026

Free Radicals in Chemical Biology: from Chemical Behavior to Biomarker Development

Free Radicals in Chemical Biology: from Chemical Behavior to Biomarker Development

Published on: April 15, 2013

New Methods to Study Gustatory Coding

New Methods to Study Gustatory Coding

Published on: June 29, 2017

Co-immunoprecipitation Assay for Studying Functional Interactions Between Receptors and Enzymes

Co-immunoprecipitation Assay for Studying Functional Interactions Between Receptors and Enzymes

Published on: September 28, 2018

Area of Science:

Biochemistry
Molecular Biology
Enzymology

Background:

Diphthamide is a unique posttranslational modification found on translation elongation factor 2 (EF2) in archaea and eukaryotes.
Its biosynthesis involves a proposed four-step pathway, with the initial step catalyzed by specialized radical S-adenosylmethionine (SAM) enzymes.

Purpose of the Study:

To investigate the unique radical SAM enzymes responsible for the first step of diphthamide biosynthesis.
To develop and apply methods for the preparation, characterization, and activity assays of these novel enzymes.

Main Methods:

Preparation and purification of unique radical SAM enzymes.
Characterization of enzyme properties and cofactor requirements.
Development and execution of activity assays to monitor enzymatic function.

Main Results:

Demonstrated the unique mechanism of diphthamide-related radical SAM enzymes in cleaving the C-S bond of SAM.
Generated and characterized active forms of the enzymes.
Established functional assays for studying enzyme kinetics and substrate interactions.

Conclusions:

The study provides crucial insights into the unique radical SAM enzymes involved in diphthamide biosynthesis.
Developed methodologies enable further investigation into the catalytic mechanisms and biological roles of these enzymes in EF2 modification.