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Related Concept Videos

Predicting Products: SN1 vs. SN202:27

Predicting Products: SN1 vs. SN2

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Nucleophilic substitution reactions of alkyl halides can proceed via an SN1 or an SN2 mechanism. While in SN2 reactions, the nucleophile attacks the substrate simultaneously as the leaving group departs, in SN1 reactions, the substrate first dissociates to give the carbocation intermediate. Various factors such as the structure of the substrate, the strength of the nucleophile, and the nature of the solvent promote one mechanism over the other.
With increased substitution on the alkyl halide,...
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Conserved Binding Sites01:49

Conserved Binding Sites

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Many proteins’ biological role depends on their interactions with their ligands, small molecules that bind to specific locations on the protein known as ligand-binding sites. Ligand-binding sites are often conserved among homologous proteins as these sites are critical for protein function.
Binding sites are often located in large pockets, and if their location on a protein’s surface is unknown, it can be predicted using various approaches. The energetic method computationally...
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Autophagy01:27

Autophagy

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Autophagy is a self-digesting process by which a cell protects itself from threats both within and outside the cell, ranging from abnormal proteins to invading bacteria. In this process, obsolete components of the cell and invading microbes are degraded by hydrolytic enzymes active in an acidic environment of the lysosomal lumen.
An autophagic pathway consists of a series of signaling events activated in response to diverse stress and physiological conditions such as food deprivation,...
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The Equilibrium Binding Constant and Binding Strength02:18

The Equilibrium Binding Constant and Binding Strength

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The equilibrium binding constant (Kb) quantifies the strength of a protein-ligand interaction. Kb can be calculated as follows when the reaction is at equilibrium:
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1° Amines to Diazonium or Aryldiazonium Salts: Diazotization with NaNO2 Overview01:26

1° Amines to Diazonium or Aryldiazonium Salts: Diazotization with NaNO2 Overview

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Nitrous acid and nitric acids are two types of acids containing nitrogen, among which nitrous acid is weaker than nitric acid. Nitrous acid with a pKa value of 3.37 ionizes in water to give a nitrite ion and the hydronium ion.
The nitrous acid is unstable. Hence, it is formed in situ from a solution of sodium nitrite and cold aqueous acids such as hydrochloric or sulfuric acid. In an acidic solution, the –OH group of nitrous acid undergoes protonation to give oxonium ion, followed by...
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1° Amines to Diazonium or Aryldiazonium Salts: Diazotization with NaNO2 Mechanism01:37

1° Amines to Diazonium or Aryldiazonium Salts: Diazotization with NaNO2 Mechanism

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Nitrous acid is a relatively weak and unstable acid prepared in situ by the reaction of sodium nitrite and cold, dilute hydrochloric acid. In an acidic solution, the nitrous acid undergoes protonation when it loses water to form a nitrosonium ion—an electrophile. Nitrous acid reacts with primary amines to give diazonium salts. The reaction is called diazotization of primary amines.
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Related Experiment Video

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Examining BCL-2 Family Function with Large Unilamellar Vesicles
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High-Throughput Screens To Identify Autophagy Inducers That Function by Disrupting Beclin 1/Bcl-2 Binding.

Wei-Chung Chiang1, Yongjie Wei1,2, Yi-Chun Kuo3

  • 1Center for Autophagy Research, Department of Internal Medicine , University of Texas Southwestern Medical Center , Dallas , Texas 75390 , United States.

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Researchers developed new assays to find compounds that stimulate autophagy, a key cellular process. They identified three novel compounds that disrupt Bcl-2 and Beclin 1 binding, potentially leading to new treatments for diseases and aging.

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Area of Science:

  • Cell Biology
  • Biochemistry
  • Pharmacology

Background:

  • Autophagy is a vital cellular process involved in homeostasis, immunity, and aging.
  • Pharmacological induction of autophagy offers potential therapeutic benefits for various diseases and aging.
  • Targeting the interaction between Bcl-2 and Beclin 1 is a key strategy for modulating autophagy.

Purpose of the Study:

  • To develop novel assays for identifying compounds that specifically disrupt Bcl-2/Beclin 1 binding.
  • To screen chemical libraries for small molecules that inhibit Bcl-2/Beclin 1 interaction and induce autophagy.
  • To identify lead compounds for developing clinically useful autophagy-inducing agents.

Main Methods:

  • Developed and utilized split-luciferase and AlphaLISA assays to measure Beclin 1/Bcl-2 binding.
  • Screened approximately 300,000 compounds from two chemical libraries.
  • Assessed compound cytotoxicity and selectivity for Bcl-2/Beclin 1 interaction over other Bcl-2 family members.

Main Results:

  • Identified three novel compounds that directly inhibit Beclin 1/Bcl-2 binding with micromolar IC50 values.
  • Demonstrated that these compounds increase autophagic flux without significant cytotoxicity.
  • Showed selectivity for disrupting Bcl-2 binding to the Beclin 1 BH3 domain compared to Bax and Bim.

Conclusions:

  • Successfully established assays to identify modulators of Beclin 1/Bcl-2 interaction.
  • Identified novel small molecules as lead compounds for developing potent and selective autophagy inducers.
  • These findings pave the way for potential therapeutic strategies targeting autophagy for disease and aging.