¹⁹ F NMR Spectroscopy Tagging and Paramagnetic Relaxation Enhancement-Based Conformation Analysis of Intrinsically...

Area of Science:

• Biochemistry
• Structural Biology
• Biophysics

Background:

• Protein-protein interactions are crucial for cellular functions.
• Intrinsically disordered proteins (IDPs) play vital roles but are challenging to study structurally.
• Understanding the dynamics of protein complexes like Myc-Max is important for cancer research.

Purpose of the Study:

• To evaluate a combined 19F NMR and PRE NMR spectroscopy approach for studying protein interactions and IDP conformational changes.
• To apply this method to the Myc-Max oncogenic transcription factor complex.
• To gain insights into the differential structural dynamics of Myc-Max.

Main Methods:

• Utilized 19F NMR spectroscopy tagging of the Myc protein with a fluorinated tag.
• Employed paramagnetic relaxation enhancement (PRE) NMR spectroscopy using MTSL-tagged Max mutants.
• Measured 19F R2 relaxation rates to probe intermolecular interactions and dynamics.

Main Results:

• Successfully demonstrated the feasibility of the combined 19F NMR and PRE NMR strategy.
• Revealed novel insights into the differential structural dynamics of the Myc-Max complex when bound to DNA and breast cancer antigen 1.
• Provided detailed information on the conformational changes and interactions within the Myc-Max complex.

Conclusions:

• The combined 19F NMR and PRE NMR method is a versatile tool for investigating protein-protein interactions and conformational changes.
• This strategy offers significant advantages for structural biology and inhibitor screening.
• The findings contribute to a better understanding of the Myc-Max complex's role in oncogenesis.