Nanosecond Molecular Motion in pHP1α Liquid-Liquid Phase Separation Captured by Solid-State NMR.

The relationship among protein structure, function, and dynamics is fundamental to biological activity, particularly in more complex biomolecular systems. Solid-state and solution-state NMR techniques offer powerful means to probe these dynamics across various time scales. However, standard assumptions about molecular motion are often challenged in phase-separated systems like phosphorylated heterochromatin protein 1 alpha (pHP1α), which exhibit both solid- and solution-like characteristics. This study investigates the nanosecond molecular motions in pHP1α liquid-liquid phase separation (LLPS) using relaxation in hetNOE-filtered HSQC signals. By systematically analyzing motions captured by hetNOE-filtered HSQC and conventional HSQC, we characterize the global dynamics site-specifically in pHP1α LLPS. Our findings reveal ∼15 ns motion in the pHP1α LLPS system, suggesting the coexistence of different dynamic phases, and support previous observations on its role in chromatin organization. This work contributes to the expanding literature on phase-separated biomolecular behavior, with implications for understanding the molecular basis of chromatin compaction and genomic stability.