We measure the relaxation rates of our protein backbone, including a 1H–15N hetero NOE experiment. In my protein, there is a specific region that forms a helix (confirmed by Cα and CO chemical shifts, as well as CD data). However, when we recorded the 1H–15N hetero NOE, the helix‐forming region showed values as low as 0.6–0.7. We used a 5‐second relaxation delay and applied the same parameters to all data processing.

Has anyone else encountered a similar issue?