NSF N-terminal domain crystal structure: models of NSF function

Area of Science:

• Molecular biology
• Structural biology
• Cell biology

Background:

• N-ethylmaleimide-sensitive factor (NSF) is a crucial ATPase in eukaryotic vesicle fusion.
• NSF, with SNAP proteins, disassembles cis-SNARE complexes post-ATP hydrolysis.
• This disassembly primes SNAREs for subsequent trans-complex formation.

Purpose of the Study:

• To determine the crystal structure of the N-terminal domain of NSF (N).
• To elucidate the structural basis for NSF's role in SNARE complex disassembly.
• To propose a mechanism for NSF-mediated conformational changes.

Main Methods:

• X-ray crystallography was used to determine the structure of the NSF N-terminal domain (N).
• Structural analysis was performed to identify key features and potential interaction sites.
• Comparative structural analysis was conducted with other known proteins.

Main Results:

• The crystal structure of the NSF N-terminal domain (N) was resolved to 1.9 Å resolution.
• The N domain comprises two subdomains forming a groove, a potential SNAP interaction site.
• Unexpected structural similarity was found between NSF N subdomains and EF-Tu domains.

Conclusions:

• The determined structure provides insights into NSF's interaction with SNAP proteins.
• The structural similarity to EF-Tu suggests a conserved mechanism for conformational changes.
• A model for a large conformational change in NSF, driving SNARE complex disassembly, is proposed.