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Related Concept Videos

Cooperative Allosteric Transitions01:58

Cooperative Allosteric Transitions

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Cooperative allosteric transitions can occur in multimeric proteins, where each subunit of the protein has its own ligand-binding site. When a ligand binds to any of these subunits, it triggers a conformational change that affects the binding sites in the other subunits; this can change the affinity of the other sites for their respective ligands. The ability of the protein to change the shape of its binding site is attributed to the presence of a mix of flexible and stable segments in the...
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Allosteric Proteins-ATCase01:19

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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.
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...
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Cofactors and Coenzymes01:24

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Enzymes are proteins made of amino acids. The functional group of each constituent amino acid catalyzes a wide variety of chemical reactions via ionic interactions or acid-base reactions. However, amino acids cannot catalyze oxidation-reduction and group transfer reactions and need to be aided by non-protein components called cofactors. Cofactors are also referred to as the chemical teeth of an enzyme.
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Cofactors and Coenzymes01:27

Cofactors and Coenzymes

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Enzymes require additional components for proper function. There are two such classes of molecules: cofactors and coenzymes. Cofactors are metallic ions and coenzymes are non-protein organic molecules. Both of these types of helper molecule can be tightly bound to the enzyme or bound only when the substrate binds.
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Structure and Coordination Determination of Peptide-metal Complexes Using 1D and 2D 1H NMR
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An evolutionary path to altered cofactor specificity in a metalloenzyme.

Anna Barwinska-Sendra1, Yuritzi M Garcia2, Kacper M Sendra1

  • 1Institute for Cell and Molecular Biosciences, Faculty of Medical Sciences, Newcastle University, Newcastle upon Tyne, NE2 4HH, UK.

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Researchers identified two key positions in metalloenzymes that control metal specificity, impacting bacterial resistance. These findings advance understanding of metalloenzyme evolution and cofactor utilization.

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Area of Science:

  • Biochemistry
  • Enzymology
  • Structural Biology

Background:

  • Metalloenzymes, crucial for biological processes, often use metal cofactors, but the determinants of metal specificity are largely unknown.
  • This lack of understanding hinders the targeted engineering of metalloenzymes for industrial and therapeutic applications.
  • The iron/manganese superoxide dismutase (SOD) family serves as a model system where metal usage cannot be predicted.

Purpose of the Study:

  • To elucidate the structural and biochemical basis of metal specificity in the superoxide dismutase (SOD) enzyme family.
  • To identify specific amino acid residues responsible for dictating cofactor metal utilization.
  • To investigate the functional consequences of altered metal specificity on enzyme activity and bacterial physiology.

Main Methods:

  • Comparative biochemical and structural analysis of two related SODs from Staphylococcus aureus with differing metal specificities.
  • Paramagnetic studies to probe the metal's environment and redox properties.
  • Site-directed mutagenesis to alter identified residues and assess the impact on metal specificity and enzyme function.
  • In vivo assays evaluating bacterial resistance to superoxide stress under metal-limited conditions.

Main Results:

  • Two specific amino acid positions were identified as critical determinants of metal specificity in SODs.
  • These residues do not directly coordinate the metal cofactor but influence its redox properties.
  • Mutating these positions significantly altered the enzyme's preferred metal cofactor.
  • The engineered mutations affected Staphylococcus aureus's ability to withstand oxidative stress when starved of essential metals.

Conclusions:

  • Subtle changes in enzyme architecture can profoundly alter metal cofactor utilization and specificity.
  • Understanding these determinants is key to engineering metalloenzymes with novel cofactor specificities.
  • These findings provide insights into the evolutionary mechanisms driving metalloenzyme adaptation and cofactor diversification.