Structure of the apoptotic protease-activating factor 1 bound to ADP
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View abstract on PubMed
Summary
This summary is machine-generated.The apoptotic protease-activating factor 1 (Apaf-1) protein remains inactive until it binds ATP, forming a complex that activates caspase-9. This study reveals the crystal structure of inactive Apaf-1, detailing how ADP locks it in a non-functional state.
Area Of Science
- Molecular Biology
- Cell Biology
- Biochemistry
Background
- Apoptosis, or programmed cell death, is crucial for development and tissue homeostasis.
- Caspases are key executioners of apoptosis, activated by specific stimuli.
- Apoptotic protease-activating factor 1 (Apaf-1) initiates caspase activation downstream of mitochondrial pathways.
Purpose Of The Study
- To elucidate the molecular mechanisms governing Apaf-1's inactive state.
- To determine the structural basis for Apaf-1's regulation before apoptosome formation.
- To understand how nucleotide binding regulates Apaf-1 function in apoptosis.
Main Methods
- X-ray crystallography of ADP-bound, WD40-deleted Apaf-1 at 2.2-Å resolution.
- Structural analysis of protein-protein interactions and nucleotide binding sites.
- Biochemical assays to study Apaf-1's interaction with nucleotides and caspase-9.
Main Results
- The crystal structure reveals how Apaf-1 maintains an inactive conformation through domain packing.
- An ADP molecule is centrally located, stabilizing interactions between four domains and burying the caspase-9 binding site.
- Apaf-1 binds and hydrolyzes ATP/dATP, suggesting nucleotide hydrolysis drives conformational changes for apoptosome assembly.
Conclusions
- The inactive state of Apaf-1 is structurally defined by specific domain interactions stabilized by ADP.
- Nucleotide binding and hydrolysis are critical for conformational changes leading to apoptosome formation.
- Understanding Apaf-1 regulation provides insights into controlling apoptosis and related diseases.
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