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Related Concept Videos

Enzymes and Activation Energy01:13

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The activation energy (or free energy of activation), abbreviated as Ea, is the small amount of energy input necessary for all chemical reactions to occur. During chemical reactions, certain chemical bonds break, and new ones form. For example, when a glucose molecule breaks down, bonds between the molecule's carbon atoms break. Since these are energy-storing bonds, they release energy when broken. However, the molecule must be somewhat contorted to get into a state that allows the bonds to...
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For many years, scientists thought that enzyme-substrate binding took place in a simple "lock-and-key" fashion. This model stated that the enzyme and substrate fit together perfectly in one instantaneous step. However, current research supports a more refined view scientists call induced fit. The induced-fit model expands upon the lock-and-key model by describing a more dynamic interaction between enzyme and substrate. As the enzyme and substrate come together, their interaction causes...
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Enzymes02:34

Enzymes

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Inside living organisms, enzymes act as catalysts for many biochemical reactions involved in cellular metabolism. The role of enzymes is to reduce the activation energies of biochemical reactions by forming complexes with its substrates. The lowering of activation energies favor an increase in the rates of biochemical reactions.
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Ketones with α protons are deprotonated by strong bases like lithium diisopropylamide (LDA) to form enolate ions. The anion is stabilized by resonance, and its hybrid structure exhibits negative charges on the carbonyl oxygen and the α carbon. This ambident nucleophile can attack an electrophile via two possible sites: the carbonyl oxygen, known as O-attack, or the α carbon, known as C-attack. The nucleophilic attack via the carbanionic site is preferred. This is due to the...
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Alkenes can be dihydroxylated using potassium permanganate.  The method encompasses the reaction of an alkene with a cold, dilute solution of potassium permanganate under basic conditions to form a cis-diol along with a brown precipitate of manganese dioxide.
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Engineering potassium activation into biosynthetic thiolase.

Andrew C Marshall1,2, John B Bruning2

  • 1School of Molecular Sciences, The University of Western Australia, Crawley, Western Australia 6009, Australia.

The Biochemical Journal
|August 2, 2021
PubMed
Summary
This summary is machine-generated.

Researchers engineered potassium ion (K+) activation into a K+-independent enzyme, demonstrating protein malleability. Specific residues near the binding site control K+ activation, offering insights into enzyme regulation.

Keywords:
acetyltransferasesenzyme activationmetalloenzymesmolecular mechanismsprotein engineering

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Area of Science:

  • Biochemistry
  • Structural Biology
  • Enzymology

Background:

  • Monovalent cations (M+) activate numerous enzymes, yet structural details are scarce.
  • Thiolases are conserved enzymes with both K+-activated and K+-independent forms.

Purpose of the Study:

  • To engineer K+-activation into a K+-independent thiolase using a structure-based approach.
  • To elucidate the molecular determinants of K+-ion allosteric regulation in enzymes.

Main Methods:

  • Structure-based protein engineering of a K+-independent thiolase.
  • Biochemical assays to assess enzyme activity and K+-dependence.
  • Bioinformatic analysis of conserved residues and evolutionary signatures.

Main Results:

  • Successfully engineered K+-activation into a K+-independent thiolase, a first demonstration of its kind.
  • Identified two key residues (tyrosine and glutamate) near the active site essential for K+ coordination and activation.
  • A distal residue and a water molecule were found to mediate substrate binding via hydrogen bonds, influencing K+-activation.

Conclusions:

  • Protein structure can be modified to introduce allosteric regulation by monovalent cations.
  • Specific amino acid residues dictate K+-ion binding and subsequent enzyme activation.
  • Evolutionary conservation patterns can predict K+-activation in enzyme families.