Improvement in protein HSQC spectra from addition of betaine

• DOI: 10.1007/s10858-025-00463-0
• Publication Date: 2025-03-10T04:39:17Z
• Authors: Finn O’Dea; Aiden J. Seargeant; Jessica Hurcum; Rodolpho do Aido-...; Michelle L. Rowe; Nicola J. Baxter; Jon P. Waltho; Jon R. Sayers; Mike P. Williamson
• Keywords: Betaine; Amide

Abstract Addition of glycine betaine up to 1 M gave rise to increased intensity for some weak signals in the HSQC spectra of barnase and Plasmodium falciparum flap endonuclease. The signals that were enhanced were low intensity signals, often from amide groups with rapid internal motion (low order parameter). The majority of signals are however somewhat weaker because of the increased viscosity. Addition of betaine is shown to cause a small but significant overall increase in order parameter, no consistent effect on conformational change on the µs-ms timescale, and a reduction in amide exchange rates, by a factor of about 3. The results are consistent with a model in which betaine leads to a reduction in fluctuations within the bulk water, which in turn produces a reduction in internal fluctuations of the protein.