Fig. 4.

¹H-¹⁵N HSQC spectra of NESG targets recorded with a 1.7-mm micro-cryoprobe at 20°C labeled with corresponding NESG target IDs. a NR26 scored as “poor” due to low sensitivity and low peak count. b HR5272A scored as “unfolded”, as all backbone amides are in the random coil region (∼7–9 ppm), and the side-chain amide resonances are largely overlapped. c GmR58A scored as “promising”, as the spectrum shows the simultaneous presence of folded regions and unfolded segments with higher peak intensity. d NsR431C scored as “good”, with uniform intensity, good dispersion, and correct peak count. In our pipeline, target GmR58A is a candidate for construct optimization using HDX-MS, while target NsR431C was promoted for uniform ¹⁵N,¹³C-enrichment and structure determination