Enzymes and Kinetics

May 28, 2024

Enzymes and Kinetics Lecture Notes

Introduction to Enzymes

  • Definition: Enzymes are biological catalysts that do not change the thermodynamics (enthalpy or Gibbs free energy) but speed up reactions.
  • Role: They help reactions proceed faster by affecting the kinetics.

Classification of Enzymes

  1. Oxidor Reductase
    • Catalyze oxidation reactions by transferring electrons.
    • Example: Alcohol dehydrogenases (convert alcohols to aldehydes or ketones).
  2. Transferases
    • Catalyze the transfer of functional groups between molecules.
    • Example: Aminotransferases (degrade amino acids by removing amino groups).
  3. Hydrolases
    • Catalyze hydrolysis (breaking bonds with water).
  4. Liases
    • Catalyze the breakdown of chemical bonds, forming new double bonds or rings.
    • Example: Pyruvate decarboxylase (removes carbon dioxide from pyruvate).
  5. Isomerases
    • Catalyze structural shifts within a molecule.
    • Example: Ribulose phosphate epimerase (converts ribulose biphosphate to xylose 5-phosphate).
  6. Ligase/Synthetases
    • Catalyze the joining of two molecules by forming new bonds.

How Enzymes Work

  • Mechanism: Lower the activation energy of a reaction by providing favorable environments, stabilizing transition states, or bringing reactive groups closer.
  • Protein Nature: All enzymes are proteins, but not all proteins are enzymes.
  • Specificity: Enzymes are highly specific, only binding to specific substrates that fit their active sites.
  • Enzyme-Substrate Complex: Formation of this complex lowers activation energy, catalyzing the reaction.
  • Regulation: Enzymes can be regulated by other molecules (activators or inhibitors) and post-translational modifications (e.g., phosphorylation).

Theories of Enzyme-Substrate Interaction

  1. Lock and Key Theory
    • Active site is already in the right conformation for the substrate.
  2. Induced Fit Model
    • Both enzyme and substrate undergo conformational changes to fully interact.

Enzyme Kinetics

  • Relation to Concentration: The rate of reaction depends on concentrations of both the enzyme and the substrate.
  • Michaelis-Menten Equation: Describes reaction rates.
    • Equation:

                 V = (Vmax * [S]) / (Km + [S])
      
  • Key Kinetic Terms:
    • Vmax: Maximum reaction rate.
    • Km: Substrate concentration at which half of the enzyme active sites are occupied.

Michaelis-Menten Curve

  • X-axis: Substrate concentration.
  • Y-axis: Reaction rate (velocity).
  • Vmax: Plateau where increasing substrate does not increase reaction rate.
  • Km: Substrate concentration at 1/2 Vmax.

Lineweaver-Burk Plot

  • Plot: Double reciprocal of Michaelis-Menten ( extV vs 1/[S]).
  • Y-intercept: 1/Vmax.
  • X-intercept: -1/Km.
  • Slope: Km/Vmax.

Effects of Local Conditions on Enzyme Activity

  1. Temperature: Reaction rate doubles for every 10°C increase until the optimal temperature, beyond which denaturation occurs.
  2. pH: Optimal pH for human enzymes is 7.4. Changes can lead to denaturation.
  3. Salinity: Affect enzyme activities by disrupting bonds, leading to conformational changes.

Regulation of Enzyme Activity

  • Feedback Regulation: Allows control of enzyme activity to maintain homeostasis.
  • Feedback Inhibition:
    • Irreversible Inhibitors: Form covalent or tight bonds with the enzyme.
    • Reversible Inhibitors:
      • Competitive: Inhibitors compete with the substrate for the active site. Can be overcome by increasing substrate concentration. Increases Km, no effect on Vmax.
      • Non-competitive: Bind to an allosteric site, changing enzyme conformation. Decreases Vmax, no effect on Km.
      • Uncompetitive: Bind only to enzyme-substrate complexes, locking substrate in place. Decrease both Vmax and Km.
  • Other Regulation Mechanisms:
    • Allosteric Activation: Enhances enzyme efficiency.
    • Phosphorylation: Covalent modifications altering enzyme activity.
    • Zymogens: Inactive enzyme forms activated by cleavage.

Conclusion

  • Enzymes play a crucial role in metabolic processes by speeding up reactions and ensuring efficiency in cellular functions.
  • Understanding enzyme kinetics and regulation is key to studying their function and impact.