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Related Concept Videos

Aldehydes and Ketones to Alkenes: Wittig Reaction Overview01:19

Aldehydes and Ketones to Alkenes: Wittig Reaction Overview

The Wittig reaction is the conversion of carbonyl compounds-aldehydes and ketones-to alkenes using phosphorus ylides, or the Wittig reagent. The reaction was pioneered by Prof. Georg Wittig, for which he was awarded the Nobel Prize in Chemistry.
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Covalently Linked Protein Regulators

Proteins can undergo many types of post-translational modifications, often in response to changes in their environment. These modifications play an important role in the function and stability of these proteins. Covalently linked molecules include functional groups, such as methyl, acetyl, and phosphate groups, and also small proteins, such as ubiquitin. There are around 200 different types of covalent regulators that have been identified.
These groups modify specific amino acids in a protein.
Aldehydes and Ketones to Alkenes: Wittig Reaction Mechanism01:14

Aldehydes and Ketones to Alkenes: Wittig Reaction Mechanism

The Wittig reaction, which converts aldehydes or ketones to alkenes using phosphorus ylides, proceeds through a nucleophilic addition‒elimination process.
The reaction begins with the nucleophilic addition between a phosphorus ylide and the carbonyl compound. Due to its carbanionic character, phosphorus ylide acts as a strong nucleophile and attacks the electrophilic carbonyl group. This generates a charge-separated dipolar intermediate called betaine. The negatively charged oxygen atom and...
Protein Modifications in the RER01:26

Protein Modifications in the RER

Modification of secretory and transmembrane proteins entering the rough ER begins in the ER lumen. These modifications aid in protein folding and stabilize the acquired tertiary structure. Protein modifications in the rough ER co-occur at different stages of protein folding.
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Conservative Site-specific Recombination and Phase Variation

Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
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Ligand Binding and Linkage00:49

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Allosteric proteins have more than one ligand binding site; the binding of a ligand to any of these sites influences the binding of ligands to the other sites. When a protein is allosteric, its binding sites are called coupled or linked.  In the case of enzymes, the site that binds to the substrate is known as the active site and the other site is known as the regulatory site. When a ligand binds to the regulatory site, this leads to conformational changes in the protein that can influence the...

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Enabling Wittig reaction on site-specific protein modification.

Ming-Jie Han1, De-Cai Xiong, Xin-Shan Ye

  • 1State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, Xue Yuan Road No. 38, Beijing 100191, China.

Chemical Communications (Cambridge, England)
|October 9, 2012
PubMed
Summary

Researchers developed an efficient aqueous Wittig reaction for protein bioconjugation. This method allows site-specific modification of proteins, enabling the introduction of diverse functional groups.

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Area of Science:

  • Bioconjugation Chemistry
  • Protein Modification
  • Organic Synthesis

Background:

  • The Wittig reaction is a fundamental organic transformation for forming carbon-carbon double bonds.
  • Protein bioconjugation is crucial for developing diagnostics, therapeutics, and biochemical tools.
  • Existing bioconjugation methods often face limitations in aqueous environments or site-specificity.

Purpose of the Study:

  • To establish an efficient and site-specific Wittig reaction in an aqueous environment for protein modification.
  • To demonstrate the applicability of this method across various molecular scales, including small molecules, peptides, and proteins.
  • To showcase the versatility of introducing diverse functional groups onto proteins.

Main Methods:

  • Investigated the Wittig reaction in aqueous buffer conditions.
  • Developed and utilized an "aldehyde tag" for site-specific protein functionalization.
  • Validated the reaction's efficiency and scope using small molecules, synthetic peptides, and recombinant proteins.

Main Results:

  • Achieved the first efficient aqueous Wittig reaction for protein bioconjugation.
  • Demonstrated high site-specificity using the aldehyde tag strategy.
  • Successfully introduced a variety of functional groups onto proteins via this method.

Conclusions:

  • The developed aqueous Wittig reaction provides a novel and efficient tool for site-specific protein bioconjugation.
  • This methodology expands the possibilities for protein engineering and functionalization in biological settings.
  • The ability to introduce diverse functionalities opens new avenues for protein-based applications.