1H, 13C, 15N backbone and side chain resonance assignment of the HNH nuclease from Streptococcus pyogenes CRISPR-Cas9

• DOI: 10.1007/s12104-019-09907-9
• Publication Date: 2019-08-03T06:13:55Z
• Authors: Helen B. Belato; Kyle W. East; George P. Lisi
• Keywords: 0301 basic medicine; 03 medical and health sciences; Streptococcus pyogenes; CRISPR-Associated Protein 9; Nuclear Magnetic Resonance, Biomolecular

HNH is one of two endonuclease domains of the clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein Cas9 that perform site-specific cleavage of double-stranded DNA. We engineered a novel construct of this critical nuclease from Streptococcus pyogenes Cas9 that not only maintains the wild-type amino acid sequence and fold, but displays enhanced thermostability when compared to the full-length Cas9 enzyme. Here, we report backbone and side chain assignments of the HNH nuclease as a foundational step toward the characterization of protein dynamics and allostery in CRISPR-Cas9.