Protein Folding

Chaperone-Assisted Protein Folding

Summary

Molecular chaperones ensure proteins achieve their functional 3D conformations without aggregating. Key systems include Hsp70, Chaperonins (GroEL-GroES), and Hsp90, operating via ATP-dependent cycles that rescue misfolded intermediates.

Key Points

  • 1Hsp70 binds hydrophobic regions to prevent aggregation
  • 2Chaperonins provide isolated folding chambers
  • 3ATP hydrolysis powers conformational cycles
  • 4Chaperone failure linked to neurodegenerative diseases

While the amino acid sequence contains all the information needed for folding (Anfinsen's dogma), the crowded cellular environment necessitates molecular chaperones to ensure proper protein folding.

Why Chaperones Are Necessary

The cellular environment presents challenges not found in test tube refolding experiments:

- Macromolecular crowding: Protein concentrations of 300-400 mg/mL

- Nascent chain vulnerability: Incomplete proteins cannot fold properly

- Aggregation risk: Exposed hydrophobic surfaces promote intermolecular interactions

Major Chaperone Systems

Hsp70 (DnaK in bacteria)

The Hsp70 system is the most versatile chaperone:

- Binds short hydrophobic peptide segments on nascent or misfolded proteins

  • Operates via ATP-dependent cycles of binding and release
  • Co-chaperones (Hsp40/DnaJ) deliver substrates and stimulate ATP hydrolysis
  • Prevents premature folding and aggregation
  • Chaperonins (GroEL-GroES / TRiC)

    Chaperonins provide a protected folding environment:

    - GroEL: Double-ring structure forming a central cavity

    - GroES: Lid that caps the folding chamber

    - Creates a hydrophilic isolation chamber where proteins can fold in isolation

  • ATP hydrolysis drives conformational cycles
  • Critical for folding ~10-15% of E. coli proteins
  • Hsp90

    Hsp90 specializes in signaling proteins:

  • Stabilizes kinases, steroid hormone receptors, and transcription factors
  • Works late in the folding pathway on near-native conformations
  • Forms a sophisticated complex with multiple co-chaperones
  • How Chaperones Work

    Chaperones don't provide folding instructions. Instead, they:

    1. Shield hydrophobic surfaces from aggregation

    2. Provide isolated folding environments (chaperonins)

    3. Unfold kinetically trapped intermediates using ATP

    4. Smooth the energy landscape toward productive folding

    Chaperones and Disease

    Failure of chaperone systems is implicated in proteopathies:

  • Alzheimer's disease
  • Parkinson's disease
  • Huntington's disease
  • Amyotrophic lateral sclerosis (ALS)