NOMO architecture is shaped by the Ig 1/10/11 interface. (A) SEC-MALS profile of FL WT NOMO (NOMO1^WT-FLAG) on a Superose 6 column. The dashed line is the UV trace, molar mass represents the total molar mass of the PDC, and protein molar mass is the corrected molar mass to remove contribution from detergent. Inset, SDS-PAGE/Coomassie staining of the NOMO1^WT-FLAG fraction obtained from preparative SEC. (B) SEC-MALS profile of the 4-mutant interface NOMO1 mutant, NOMO^4Mut-FLAG, insert as in A. (C and D) Model representation drawn from the results shown in A and B. (E) Far-UV circular dichroism spectra of WT (blue) and 4-Mut (red) NOMO LD in the native conformation, and 6 M guanidine hydrochloride (GuHCl) treatment to induce unfolding (WT, tan; 4-mutant, light blue), as indicated by the loss of secondary structure. Curves represent normalized mean ± SD of n = 3 replicates. SEC-MALS, SEC linked to multi-angle light scattering. PDC, protein detergent complex. Source data are available for this figure: SourceData F4.