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Related Experiment Videos

Proteasomes and their associated ATPases: a destructive combination.

David M Smith1, Nadia Benaroudj, Alfred Goldberg

  • 1Department of Cell Biology, Harvard Medical School, 240 Longwood Avenue, Boston, MA 02115, USA.

Journal of Structural Biology
|August 22, 2006
PubMed
Summary

Protein degradation requires ATP hydrolysis by proteasome-associated AAA ATPases like PAN. ATP binding, not hydrolysis, facilitates substrate translocation after unfolding, revealing ATP

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Structural Biology

Background:

  • Protein degradation is essential for cellular function and regulated by the proteasome.
  • ATP hydrolysis by AAA ATPases is crucial for proteasome-mediated protein degradation.
  • The archaeal proteasome-activating nucleotidase (PAN) complex serves as a model for understanding ATPase function in proteolysis.

Purpose of the Study:

  • To review recent insights into the multistep mechanisms of AAA ATPases associated with proteasomes.
  • To elucidate the specific roles of ATP binding and hydrolysis in protein degradation.
  • To highlight the advantages of studying the PAN complex for mechanistic and structural insights.

Main Methods:

  • Single-particle electron microscopy (EM) to determine the structure of the PAN complex.

Related Experiment Videos

  • Biochemical assays to investigate the roles of ATP binding and hydrolysis.
  • Comparative analysis with the eukaryotic 26S proteasome.

    • Main Results:

    • The PAN complex caps the 20S proteasome, resembling densities in the eukaryotic 19S regulatory complex.
    • ATP binding promotes PAN-20S complex formation and opens a gate for substrate entry.
    • Substrate translocation into the 20S proteasome requires ATP binding but not hydrolysis, occurring via facilitated diffusion.
    • ATP hydrolysis is essential only for the unfolding of globular proteins.

    Conclusions:

    • ATP plays multiple roles in proteolysis, with distinct requirements for binding and hydrolysis.
    • The PAN complex provides a simplified model for understanding the mechanisms of AAA ATPases in proteasome function.
    • The 26S proteasome likely employs similar ATP-dependent mechanisms for protein degradation.