Unique properties of Zika NS2B-NS3pro complexes as decoded by experiments and MD simulations

Amrita Roy; Liangzhong Lim; Shagun Srivastava; Jianxing Song

Biophysical characterization-We acquired 1 H NMR one-dimensional spectra for both linked and unlinked NS2B-NS3pro ( Fig. 2A) . Both spectra have similar up-field peaks manifested, suggesting that both of them are folded (12, 20, 23, 24) . However, it is interesting to note that the peaks of the linked complex are much more broad than those of the unlinked, implying that the linkage may introduce µs-ms conformational dynamics, as previously observed on the linked Dengue NS2B-NS3pro (12, 20, 23) . Indeed, we attempted to acquire 1 H-15 N NMR HSQC spectrum of the linked protease, but failed to detect any peaks due to significant peak broadening. By contrast, despite having almost the same sequences, 15 N-labeled Zika NS3pro in the unlinked complex has a well-dispersed HSQC spectrum, indicating that the NS3pro is well-folded, although many HSQC peaks were still too broad to be detectable (Fig. 2B ).