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Crystal structure of Mycobacterium tuberculosis ClpP1P2 suggests a model for peptidase activation by AAA+ partner
Karl R Schmitz1, Daniel W Carney2, Jason K Sello2
1Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02139; and.
Abstract:
Caseinolytic peptidase P (ClpP), a double-ring peptidase with 14 subunits, collaborates with ATPases associated with diverse activities (AAA+) partners to execute ATP-dependent protein degradation. Although many ClpP enzymes self-assemble into catalytically active homo-tetradecamers able to cleave small peptides, the Mycobacterium tuberculosis enzyme consists of discrete ClpP1 and ClpP2 heptamers that require a AAA+ partner and protein-substrate delivery or a peptide agonist to stabilize assembly of the active tetradecamer. Here, we show that cyclic acyldepsipeptides (ADEPs) and agonist peptides synergistically activate ClpP1P2 by mimicking AAA+ partners and substrates, respectively, and determine the structure of the activated complex. Our studies establish the basis of heteromeric ClpP1P2 assembly and function, reveal tight coupling between the conformations of each ring, show that ADEPs bind only to one ring but appear to open the axial pores of both rings, provide a foundation for rational drug development, and suggest strategies for studying the roles of individual ClpP1 and ClpP2 rings in Clp-family proteolysis.
Insights
Mycobacterium tuberculosis ClpP1P2 proteases require activators to form functional complexes. Cyclic acyldepsipeptides (ADEPs) and agonist peptides synergistically activate ClpP1P2, mimicking AAA+ partners and substrates for protein degradation.
Area of Science:
- Biochemistry
- Molecular Biology
- Structural Biology
Background:
- Caseinolytic peptidase P (ClpP) is a crucial protease involved in ATP-dependent protein degradation.
- Unlike many ClpP enzymes, Mycobacterium tuberculosis ClpP1P2 forms discrete heptamers requiring partners for activation.
Purpose of the Study:
- To elucidate the mechanism of activation and structure of the Mycobacterium tuberculosis ClpP1P2 protease complex.
- To investigate the synergistic activation by cyclic acyldepsipeptides (ADEPs) and agonist peptides.
Main Methods:
- X-ray crystallography to determine the structure of the activated ClpP1P2 complex.
- Biochemical assays to study enzyme activity and activation mechanisms.
Main Results:
- ADEPs and agonist peptides synergistically activate ClpP1P2 by mimicking AAA+ partners and substrates.
- The structure of the activated ClpP1P2 complex reveals tight coupling between the conformations of the two rings.
- ADEPs bind to one ring but influence the axial pore opening of both rings.
Conclusions:
- This study establishes the basis for heteromeric ClpP1P2 assembly and function.
- The findings provide a foundation for developing novel drugs targeting Clp-family proteolysis.
- Insights into ClpP1 and ClpP2 ring roles in proteolysis are suggested.
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