Higher Contrast in ¹H-Observed NMR Ligand Screening with the PEARLScreen Experiment

Area of Science:

• Biochemistry
• Structural Biology
• Chemical Biology

Background:

• Nuclear Magnetic Resonance (NMR) is crucial for fragment-based drug design (FBDD).
• Established NMR ligand screening methods include T1ρ, waterLOGSY, and Saturation Transfer Difference (STD) spectroscopy.
• Optimizing protein concentration is key for efficient NMR-based screening.

Purpose of the Study:

• Introduce PEARLScreen, a new perfect echo-based NMR experiment.
• Demonstrate PEARLScreen's enhanced binding sensitivity for ligand screening.
• Evaluate PEARLScreen's performance across a wide range of NMR field strengths.

Main Methods:

• Developed a novel perfect echo-based NMR experiment named PEARLScreen.
• Utilized longer relaxation delays and active exchange broadening for sensitivity enhancement.
• Tested the experiment on NMR spectrometers from 80 to 1200 MHz.

Main Results:

• PEARLScreen achieved superior binding sensitivity compared to conventional NMR screening methods.
• Reduced protein concentration by up to one order of magnitude without sensitivity loss.
• At 1200 MHz, protein reduction by a factor of 40 was achievable due to enhanced exchange effects and larger compound mixtures.

Conclusions:

• PEARLScreen offers significant advantages in sensitivity and protein concentration reduction for FBDD.
• The experiment demonstrates robust performance across various NMR field strengths.
• PEARLScreen is poised to become a standard for 1H-detected ligand screening.