Self-assembly of guanosine-5′-monophosphate into G-quadruplexes induces a cation-specific (sodium vs. potassium), intense terahertz Raman optical activity signal. The supramolecular helical arrangement gives rise to a distinct chiral vibrational signature in the low-frequency region.

Raman optical activity (ROA) provides unique chiroptical insights into biomolecular structure but suffers from inherently weak signals. A substantial enhancement in the low-wavenumber/THz region (50–250 cm⁻¹/1.5–7.5 THz) of ROA spectra upon the formation of mononucleotide G-quadruplexes (mG4) from guanosine-5′-monophosphate (5′rGMP) is reported. Using concentration-dependent Raman and ROA measurements, a nearly 100-fold increase in ROA intensity upon aggregation is identified, with spectral features highly sensitive to cation-mediated structural variations. These effects can be traced to long-range chiral interactions and distinct stacking arrangements within mG4. While strong terahertz Raman optical activity (THz-ROA) signals have previously been observed in globular proteins and α-helical polypeptides, the findings of this study represent the first evidence of this phenomenon in nucleotides, demonstrating that noncovalent self-assembly into supramolecular helices can give rise to intense low-frequency chiroptical responses. This establishes THz-ROA as a powerful background-free tool for studying supramolecular chirality and nucleotide self-assembly, with potential applications in characterizing biologically relevant G-quadruplex structures.