Supplementary Figure 4: NMR signal assignments and structural analysis of ∆J∆K.
(a) Sequence and NMR-derived secondary structure of ∆J∆K. Colors indicate the sequential assignment in (c). (b) Overlay of the 1H-1H NOESY spectra of [u-2H,{H1′,H2′,H2,H8}-Ade] ∆J∆K (black) and [u-2H,{H1′,H2′,H2,H8}-Ade] J-K (gray). (c) 1H-1H NOESY spectrum of the imino proton region and imino proton walk of ∆J∆K that supports the secondary structure of ∆J∆K. The imino walks are shown with red, cyan, and purple lines, respectively. (d) Sequential walk of H1′–H6 (pyrimidine) or H1′–H8 (purine) connectivity for the assignments. Labels show the intraresidual H1′–H6 or H1′–H8 signals of each residue. Green, cyan and purple lines represent the sequential walk of the segments shown in the same colors in (a). Asterisks indicate that the signals are below the threshold. (e) 1H-1H NOESY spectrum of [u-2H, {H1′,H2′,H2,H8}-Ade] ∆J∆K in H2O. (f) Regions of 1H-1H NOESY spectra of unlabeled ∆J∆K (top) and [u-2H, 1H-Ade/Gua] ∆J∆K (bottom). (g-i) Regions of 1H-1H NOESY spectra showing the NOE connectivities from the ASL to the J and K domains. (j) A region of 1H-1H NOESY spectrum of [u-2H, {H1′,H2′,H2,H8}-Ade] ∆J∆K in H2O. The long range connectivities that shows the register of the ASL to the J and K domains are labeled in red.
