Maintenance of native-like protein dynamics may not be required for engineering functional proteins

Area of Science:

• Biochemistry and Molecular Biology
• Structural Biology
• Protein Engineering

Background:

• Protein dynamics are crucial for understanding protein function.
• Investigating the impact of laboratory engineering on protein dynamics is of general interest.
• Homologous enzymes often share conserved structural and functional properties.

Purpose of the Study:

• To investigate the impact of laboratory engineering on protein dynamics.
• To compare the dynamics of naturally evolved homologous enzymes with a laboratory-engineered chimera.
• To determine if conserved protein dynamics are essential for conserved protein function.

Main Methods:

• Comparative analysis of protein dynamics using techniques like NMR spectroscopy or molecular dynamics simulations.
• Structural and functional characterization of homologous enzymes and a laboratory-engineered chimera.
• High-resolution crystallography to assess structural integrity.

Main Results:

• Two homologous enzymes with conserved structure and function also exhibited conserved dynamics.
• A laboratory-engineered chimeric enzyme showed significantly different dynamics on the millisecond timescale.
• Despite altered dynamics, the engineered chimera retained its function and high-resolution crystal structure.
• The engineered chimera demonstrated functional tolerance to modified dynamics during catalytic turnover.

Conclusions:

• Laboratory engineering can lead to altered protein dynamics without compromising function.
• Conserved protein dynamics may not be a strict requirement for engineering functional proteins.
• Functional tolerance to dynamic variations suggests flexibility in protein engineering strategies.