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

Diels–Alder Reaction Forming Bridged Bicyclic Products: Stereochemistry01:29

Diels–Alder Reaction Forming Bridged Bicyclic Products: Stereochemistry

6.4K
Diels–Alder reactions between cyclic dienes locked in an s-cis configuration and dienophiles yield bridged bicyclic products.
6.4K
Regioselectivity and Stereochemistry of Acid-Catalyzed Hydration02:34

Regioselectivity and Stereochemistry of Acid-Catalyzed Hydration

9.9K
The rate of acid-catalyzed hydration of alkenes depends on the alkene's structure, as the presence of alkyl substituents at the double bond can significantly influence the rate.
9.9K
Intramolecular Claisen Condensation of Dicarboxylic Esters: Dieckmann Cyclization01:13

Intramolecular Claisen Condensation of Dicarboxylic Esters: Dieckmann Cyclization

3.5K
Dieckmann cyclization is an intramolecular Claisen condensation of diesters. The reaction occurs in the presence of a base and generates a cyclic β-ketoester as the final product. Commonly, 1, 6 and 1, 7-diesters are preferred substrates for the reaction since the generated five, and six-membered cyclic β-keto esters are particularly more stable.
3.5K
Alkylation of β-Diester Enolates: Malonic Ester Synthesis01:14

Alkylation of β-Diester Enolates: Malonic Ester Synthesis

4.3K
Malonic ester synthesis is a method to obtain α substituted carboxylic acids from ꞵ-diesters such as diethyl malonate and alkyl halides.
4.3K
Diels–Alder Reaction Forming Cyclic Products: Stereochemistry01:28

Diels–Alder Reaction Forming Cyclic Products: Stereochemistry

5.3K
The Diels–Alder reaction is one of the robust methods for synthesizing unsaturated six-membered rings. The reaction involves a concerted cyclic movement of six π electrons: four π electrons from the diene and two π electrons from the dienophile.
5.3K
Indirect-Acting Cholinergic Agonists: Mechanism of Action01:18

Indirect-Acting Cholinergic Agonists: Mechanism of Action

3.0K
Indirect-acting cholinergic agonists work by interacting with an enzyme called acetylcholinesterase (AChE) in the synaptic cleft. They can be reversible or irreversible inhibitors and have different effects on the enzyme.
Reversible inhibitors like edrophonium bind to a specific part of the enzyme called the anionic catalytic site. They form noncovalent bonds, which means they are not strongly attached to the enzyme. This creates a temporary and less stable enzyme–inhibitor complex,...
3.0K

You might also read

Related Articles

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

Sort by
Same author

Stabilizing Proteins by Chemical Cross-Linking: Insights into Conformation, Unfolding, and Aggregation Using Native Ion Mobility Mass Spectrometry.

Analytical chemistry·2025
Same author

Detection and characterisation of ligand-induced conformational changes in acetylcholine binding proteins using biosensors and X-ray crystallography.

RSC chemical biology·2025
Same author

Helicity-Dependent Enzymatic Peptide Cyclization.

Journal of peptide science : an official publication of the European Peptide Society·2025
Same author

Structure-Based Design of Bicyclic Helical Peptides That Target the Oncogene β-Catenin.

Angewandte Chemie (International ed. in English)·2024
Same author

Identification of an H-Ras nanocluster disrupting peptide.

Communications biology·2024
Same author

Binding Dynamics of a Stapled Peptide Targeting the Transcription Factor NF-Y.

Chembiochem : a European journal of chemical biology·2024
Same journal

A Domino-Synthesized Dicoordinate Copper(I) Bis-imidazopyridine Complex Triggering Cuproptosis/Ferroptosis for Enhanced Cancer Immunotherapy.

Angewandte Chemie (International ed. in English)·2026
Same journal

Mirror-Symmetric Organic Two-Dimensional Crystals for Alternative Photon Transport Pathways.

Angewandte Chemie (International ed. in English)·2026
Same journal

Cobalt-Catalyzed Migratory E-Selective Asymmetric Aza-Nozaki-Hiyama-Kishi Coupling.

Angewandte Chemie (International ed. in English)·2026
Same journal

Facile Synthesis of α,ω-Dihydroxy Telechelic Macromonomers From Ethylene and α-Olefins for Recyclable Alternating Block Copolymers.

Angewandte Chemie (International ed. in English)·2026
Same journal

Multi-Atom Sub-Nanometer Assemblies on Interpenetrating Multi-Chambered N/C Nanospheres.

Angewandte Chemie (International ed. in English)·2026
Same journal

A Synergistic C<sub>2+</sub> Alcohols/Olefins-Intermediated Pathway Boosts CO<sub>2</sub> Hydrogenation to Aromatics.

Angewandte Chemie (International ed. in English)·2026
See all related articles

Related Experiment Video

Updated: Mar 21, 2026

Expression, Purification, Crystallization, and Enzyme Assays of Fumarylacetoacetate Hydrolase Domain-Containing Proteins
10:21

Expression, Purification, Crystallization, and Enzyme Assays of Fumarylacetoacetate Hydrolase Domain-Containing Proteins

Published on: June 20, 2019

25.0K

Multicyclic D-Stereospecific Hydrolase Dimer With High Sustained Activity.

Anissa Haim1, Sandra Liebscher2, Rasmus Klintrot1

  • 1Department of Chemistry and Pharmaceutical Sciences, VU University Amsterdam, Amsterdam, The Netherlands.

Angewandte Chemie (International Ed. in English)
|March 20, 2026
PubMed
Summary
This summary is machine-generated.

Protein macrocyclization using INCYPRO enhances enzyme stability against heat and solvents. This chemical engineering approach created robust biocatalysts, including a multicyclic dimer with improved activity and resilience.

Keywords:
INCYPRObioconjugationcross‐linkingenzymesmacrocyclizationprotein engineering

More Related Videos

Enzymatic Cascade Reactions for the Synthesis of Chiral Amino Alcohols from L-lysine
09:14

Enzymatic Cascade Reactions for the Synthesis of Chiral Amino Alcohols from L-lysine

Published on: February 16, 2018

12.8K
Structural Biology and Analytical Chemistry Approaches for Characterizing C-Glycoside Metabolic Enzymes in Human Gut Microbiota
13:35

Structural Biology and Analytical Chemistry Approaches for Characterizing C-Glycoside Metabolic Enzymes in Human Gut Microbiota

Published on: May 23, 2025

1.1K

Related Experiment Videos

Last Updated: Mar 21, 2026

Expression, Purification, Crystallization, and Enzyme Assays of Fumarylacetoacetate Hydrolase Domain-Containing Proteins
10:21

Expression, Purification, Crystallization, and Enzyme Assays of Fumarylacetoacetate Hydrolase Domain-Containing Proteins

Published on: June 20, 2019

25.0K
Enzymatic Cascade Reactions for the Synthesis of Chiral Amino Alcohols from L-lysine
09:14

Enzymatic Cascade Reactions for the Synthesis of Chiral Amino Alcohols from L-lysine

Published on: February 16, 2018

12.8K
Structural Biology and Analytical Chemistry Approaches for Characterizing C-Glycoside Metabolic Enzymes in Human Gut Microbiota
13:35

Structural Biology and Analytical Chemistry Approaches for Characterizing C-Glycoside Metabolic Enzymes in Human Gut Microbiota

Published on: May 23, 2025

1.1K

Area of Science:

  • Biocatalysis
  • Protein Engineering
  • Structural Biology

Background:

  • Enzymes are crucial catalysts but often lack stability under industrial conditions like high temperatures or organic cosolvents.
  • Sequence-based methods improve enzyme stability, but chemical protein engineering offers alternative strategies by modifying the natural amino acid repertoire.
  • Tailoring enzyme function and robustness is essential for expanding their applications in various processes.

Purpose of the Study:

  • To improve the stability and robustness of a D-stereospecific hydrolase using chemical protein engineering.
  • To investigate the efficacy of in situ protein cyclization (INCYPRO) for enzyme stabilization.
  • To explore the potential of macrocyclization for creating enhanced biocatalysts.

Main Methods:

  • Application of the in situ cyclization of proteins (INCYPRO) technique to a D-stereospecific hydrolase.
  • Site-specific macrocyclization to enhance enzyme resilience to thermal and cosolvent stress.
  • X-ray crystallography to confirm the structure of cross-linked protein dimers.

Main Results:

  • Site-specific macrocyclization significantly improved the enzyme's resilience to heat and cosolvent stress.
  • An unexpected cross-linked protein dimer exhibited enhanced activity and thermal stability.
  • Engineered multicyclic enzyme dimers with multiple cross-linking sites retained high activity and outperformed wild-type enzymes under stress.

Conclusions:

  • Protein macrocyclization is a versatile strategy for stabilizing both monomeric and multimeric enzymes.
  • INCYPRO provides a powerful route to engineer robust biocatalysts with improved functional properties.
  • Chemical protein engineering, through macrocyclization, offers complementary approaches to enhance enzyme performance.