Mapping protein dynamics at high spatial resolution with temperature-jump X-ray crystallography.

Area of Science:

• Structural Biology
• Biophysics
• Enzymology

Background:

• Protein conformational dynamics are crucial for biological functions like enzyme catalysis.
• Time-resolved crystallography can study protein motions but requires effective perturbations.
• Understanding protein dynamics at atomic resolution remains a significant challenge.

Purpose of the Study:

• To develop and apply a novel method combining temperature jump and time-resolved crystallography.
• To visualize atomic-level protein motions and their functional relevance in lysozyme.
• To investigate how inhibitor binding affects enzyme dynamics.

Main Methods:

• Coupling solvent-based temperature jump with time-resolved crystallography.
• Utilizing lysozyme as a model dynamic enzyme.
• Applying inhibitor binding as an orthogonal perturbation.

Main Results:

• Observed widespread atomic vibrations on the nanosecond timescale.
• Identified localized structural fluctuations coupled to the active site on the submillisecond timescale.
• Demonstrated that inhibitor binding blocks motions crucial for energy dissipation and catalysis.

Conclusions:

• The coupled temperature jump and time-resolved crystallography method provides a universal approach to study protein dynamics.
• Protein motions are essential for enzyme catalysis and can be precisely controlled.
• This technique opens new avenues for understanding and engineering protein function.