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Allosteric Regulation01:08

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Allosteric regulation of enzymes occurs when the binding of an effector molecule to a site that is different from the active site causes a change in the enzymatic activity. This alternate site is called an allosteric site, and an enzyme can contain more than one of these sites. Allosteric regulation can either be positive or negative, resulting in an increase or decrease in enzyme activity. Most enzymes that display allosteric regulation are metabolic enzymes involved in the degradation or...
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Cyclic Adenosine Monophosphate (cAMP) is an essential second messenger that activates protein kinase A (PKA) and regulates various biological processes. A single epinephrine molecule binds to GPCR and activates several heterotrimeric G proteins, each stimulating multiple adenylyl cyclase, amplifying the signal, and synthesizing large numbers of cAMP molecules. Small changes in cAMP concentration affect PKA activity. The binding of four cAMP molecules induces a conformational change in PKA,...
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Binding sites linkages can regulate a protein's function.  For example, enzyme activity is often regulated through a feedback mechanism where the end product of the biochemical process serves as an inhibitor.
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Cellular needs and conditions vary from cell to cell and change within individual cells over time. For example, the required enzymes and energetic demands of stomach cells are different from those of fat storage cells, skin cells, blood cells, and nerve cells. Furthermore, a digestive cell works much harder to process and break down nutrients during the time that closely follows a meal compared with many hours after a meal. As these cellular demands and conditions vary, so do the amounts and...
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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.
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Allosteric Modulation of AMPK Enzymatic Activity: In Vitro Characterization.

J Ward1, A R Reyes1, R G Kurumbail2

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Methods in Enzymology
|March 4, 2017
PubMed
Summary
This summary is machine-generated.

AMP-activated protein kinase (AMPK) is a crucial cellular energy sensor. This study details assay methods for discovering and characterizing small-molecule AMPK activators, vital for understanding cellular metabolism and stress responses.

Keywords:
AMPK activatorAMPK in autophagyAMPK in vitro assaysEnzyme activation

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Area of Science:

  • Biochemistry
  • Cellular Biology
  • Molecular Pharmacology

Background:

  • AMP-activated protein kinase (AMPK) acts as a master regulator of cellular energy homeostasis.
  • AMPK activation is critical during cellular stress, influencing metabolism, growth, and autophagy.
  • AMPK activation involves AMP binding to the gamma subunit, enhancing phosphorylation, preventing dephosphorylation, and allosteric activation.

Purpose of the Study:

  • To describe assay formats for identifying small-molecule activators of AMPK.
  • To facilitate the characterization of compounds targeting AMPK activity.

Main Methods:

  • Development and application of various assay formats.
  • Screening for small-molecule compounds that modulate AMPK activity.

Main Results:

  • Several effective assay formats for AMPK activator discovery were established.
  • Characterization methodologies for identified compounds were outlined.

Conclusions:

  • Assays are essential for identifying and characterizing small-molecule AMPK activators.
  • Targeting AMPK with small molecules holds therapeutic potential for metabolic and stress-related disorders.