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Related Concept Videos

Caspases01:24

Caspases

Caspase, a family of cysteine proteases, serve as effectors in apoptosis. The ced3 gene in C.elegans was first identified to be involved in apoptosis. This gene encodes the ced-3 caspase that is similar to the interleukin-1-beta converting enzyme or ICE in mammals. In addition to apoptosis, caspases also function in the inflammatory response. Inflammatory caspases are essential in activating pro-inflammatory cytokines that recruit immune cells and block the replication of pathogens inside cells.
Allosteric Proteins-ATCase01:19

Allosteric Proteins-ATCase

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.
Aspartate transcarbamoylase (ATCase) is a cytosolic enzyme that catalyzes the condensation of L-aspartate and carbamoyl phosphate to  N-carbamoyl-L-aspartate. This reaction is the first step in pyrimidine biosynthesis. UTP and CTP, the end products of the pyrimidine synthesis pathway,...
The Intrinsic Apoptotic Pathway01:31

The Intrinsic Apoptotic Pathway

Internal cellular stress, such as cellular injury or hypoxia, triggers intrinsic apoptosis. The B-cell lymphoma 2 (Bcl-2) family of proteins are the primary regulators of the intrinsic apoptotic pathway. For example, during DNA damage, checkpoint proteins, such as Ataxia Telangiectasia Mutated (ATM protein) and Checkpoints Factor-2 (Chk2) proteins, are activated. These proteins phosphorylate p53 which further activates pro-apoptotic proteins, such as Bax, Bak, PUMA, and Noxa, and inhibits...
The Extrinsic Apoptotic Pathway01:17

The Extrinsic Apoptotic Pathway

The extrinsic apoptotic pathway is initiated when extracellular death-inducing signals, such as specific cytokines, activate the death receptors expressed on the cell surface. The immune cells involved in this pathway are natural killer cells (NK cells) and cytotoxic T-lymphocytes. NK cells are critical in innate immune response, while cytotoxic T-lymphocytes are associated with adaptive immune response. These cells recognize specific receptors expressed on the altered cells and activate...

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Related Experiment Video

Updated: May 22, 2026

Exploring Caspase Mutations and Post-Translational Modification by Molecular Modeling Approaches
05:56

Exploring Caspase Mutations and Post-Translational Modification by Molecular Modeling Approaches

Published on: October 13, 2022

Allosteric modulation of caspase 3 through mutagenesis.

Jad Walters1, Joshua L Schipper, Paul Swartz

  • 1Department of Molecular and Structural Biochemistry, North Carolina State University, Raleigh, NC 27695, USA.

Bioscience Reports
|May 22, 2012
PubMed
Summary

A specific mutation in caspase 3 (CASP3) enzyme blocks its activity by altering its structure. This finding suggests allosteric regulation involves a range of inactive states, not just simple on/off switches.

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Area of Science:

  • Biochemistry
  • Structural Biology
  • Enzymology

Background:

  • Caspase 3 (CASP3) is a key executioner protease in apoptosis.
  • Allosteric inhibition is a strategy to modulate enzyme activity, but its precise mechanisms in CASP3 are not fully understood.
  • Previous models proposed discrete 'on' and 'off' states for allosteric regulation.

Purpose of the Study:

  • To investigate the structural and functional consequences of a specific mutation (Val266 to Histidine) in the CASP3 allosteric site.
  • To explore the role of coupled mutations (E124A, Y197C) in modulating CASP3 activity.
  • To challenge the discrete state model of allosteric inhibition and propose a more comprehensive view.

Main Methods:

  • X-ray crystallography to determine protein structures.
  • Enzymatic activity assays to measure CASP3 function.
  • Molecular dynamics simulations to analyze protein dynamics and conformational changes.

Main Results:

  • The Val266Histidine mutation abolished CASP3 activity, consistent with models of allosteric inhibition.
  • Mutations E124A and Y197C, when coupled to His266, further influenced enzyme activity and structure.
  • Structural analysis revealed that the mutation-induced defects propagate to the active site via a surface helix.
  • Results indicate that allosteric regulation of CASP3 is more complex than a simple shift between two states.

Conclusions:

  • Allosteric regulation of CASP3 cannot be adequately described by a simple two-state (on/off) model.
  • A more accurate representation requires considering an ensemble of inactive states.
  • Subtle structural alterations can lead to the population of these inactive states, offering new insights into enzyme regulation.