Related Experiment Video

Updated: Jul 7, 2026

In vivo and in vitro Studies of Adaptor-clathrin Interaction
17:14

In vivo and in vitro Studies of Adaptor-clathrin Interaction

Published on: January 26, 2011

Distinct structural elements of the adaptor ClpS are required for regulating degradation by ClpAP

Jennifer Y Hou1, Robert T Sauer, Tania A Baker

  • 1Department of Biology, 68-523, 77 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA.

Nature Structural & Molecular Biology
|February 26, 2008
PubMed

Insights

The adaptor protein ClpS controls which proteins the ClpAP protease degrades. Varying ClpS amounts changes ClpAP’s substrate preference, revealing ClpS acts as an allosteric effector.

Area of Science:

  • Molecular biology
  • Protein degradation
  • Enzyme regulation

Background:

  • AAA+ ATPases are crucial molecular machines.
  • Adaptor proteins regulate substrate specificity in AAA+ ATPase complexes.
  • The Escherichia coli ClpAP protease degrades proteins, with specificity influenced by adaptors like ClpS.

Purpose of the Study:

  • To investigate how the adaptor ClpS regulates substrate selection by the ClpAP protease.
  • To elucidate the mechanisms by which ClpS controls the degradation of different protein substrates.

Main Methods:

  • Biochemical assays to measure protein degradation rates.
  • Analysis of substrate binding and delivery mechanisms.
  • Mutational analysis of the ClpS adaptor protein.

Main Results:

  • ClpS binding stoichiometry dictates substrate preference: high ClpS favors N-end-rule substrates, while low ClpS allows degradation of both N-end-rule and ssrA-tagged proteins.
  • ClpS utilizes distinct mechanisms for substrate delivery and inhibition; its N-terminal segment is key for N-end-rule substrates but not ssrA-tagged proteins.
  • ClpS does not inhibit ssrA-tagged protein degradation via steric hindrance, and substrate tethering alone doesn't explain ClpS function.

Conclusions:

  • ClpS acts as an allosteric effector of ClpAP, modulating its substrate specificity.
  • Adaptor stoichiometry is a critical factor in determining AAA+ protease substrate selection.
  • This study expands the understanding of how AAA+ adaptors control protein degradation pathways.

More Related Videos

Measuring Composition of CD95 Death-Inducing Signaling Complex and Processing of Procaspase-8 in this Complex07:17

Measuring Composition of CD95 Death-Inducing Signaling Complex and Processing of Procaspase-8 in this Complex

Published on: August 2, 2021

Related Concept Videos

Clathrin Coated Vesicles01:12

Clathrin Coated Vesicles

Clathrin-coated vesicles use endocytosis to transport receptors and lysosomal hydrolases from the Golgi to the lysosome in the late secretory pathway. Clathrin-mediated endocytosis was the first described endocytic process, and Clathrin-coated vesicles remain one of the most well-studied transport vesicles. The molecular machinery that generates clathrin-coated vesicles comprises over 50 proteins that precisely coordinate vesicle formation. Cell surface receptors concentrated in indented sites...
Regulated Protein Degradation02:58

Regulated Protein Degradation

It is vital to regulate the activity of enzymatic as well as non-enzymatic proteins inside the cell. This can be achieved either through creating a balance between their rate of synthesis and degradation or regulating the intrinsic activity of the protein. Both these regulation mechanisms play an essential role in the normal functioning of cells.
Protein degradation plays two important roles in the cells. It helps to protect cells from misfolded or damaged proteins before they lead to a...
Regulated Protein Degradation02:58

Regulated Protein Degradation

It is vital to regulate the activity of enzymatic as well as non-enzymatic proteins inside the cell. This can be achieved either through creating a balance between their rate of synthesis and degradation or regulating the intrinsic activity of the protein. Both these regulation mechanisms play an essential role in the normal functioning of cells.
Protein degradation plays two important roles in the cells. It helps to protect cells from misfolded or damaged proteins before they lead to a...
The Replisome03:01

The Replisome

DNA replication is carried out by a large complex of proteins that act in a coordinated matter to achieve high-fidelity DNA replication. Together this complex is known as the DNA replication machinery or the replisome.
The synthesis of the leading and lagging strands is a highly coordinated process. To explain this, the “Trombone model” was proposed by Bruce Alberts in 1980. The DNA loop formation starts when a primer is synthesized on the parent lagging strand. The loop grows with the...
Anaphase Promoting Complex00:50

Anaphase Promoting Complex

The stepwise destruction of specific proteins is necessary for the progression and completion of the cell cycle. Such proteins are ubiquitinated by ubiquitin ligases and then subsequently destroyed by the proteasome. The SCF (Skp1/Cullin/F-box) and the anaphase-promoting complex (APC) are two important ubiquitin ligases involved in cell cycle progression. While SCF is active throughout the cell cycle, APC gets activated during metaphase to anaphase transition. Cdc20 or Cdh1 binds to APC and...
Anaphase Promoting Complex00:50

Anaphase Promoting Complex

The stepwise destruction of specific proteins is necessary for the progression and completion of the cell cycle. Such proteins are ubiquitinated by ubiquitin ligases and then subsequently destroyed by the proteasome. The SCF (Skp1/Cullin/F-box) and the anaphase-promoting complex (APC) are two important ubiquitin ligases involved in cell cycle progression. While SCF is active throughout the cell cycle, APC gets activated during metaphase to anaphase transition. Cdc20 or Cdh1 binds to APC and...