Open Access1H, 13C, 15N backbone and side chain resonance assignment of the HNH nuclease from Streptococcus pyogenes CRISPR-Cas9
Author(s) -
Helen B. Belato,
Kyle W. East,
George P. Lisi
Publication year - 2019
Publication title -
biomolecular nmr assignments
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.359
H-Index - 16
eISSN - 1874-2718
pISSN - 1874-270X
DOI - 10.1007/s12104-019-09907-9
Subject(s) - crispr , nuclease , streptococcus pyogenes , cas9 , endonuclease , cleavage (geology) , palindrome , dna , genetics , biology , side chain , chemistry , computational biology , biochemistry , gene , paleontology , organic chemistry , fracture (geology) , bacteria , staphylococcus aureus , polymer
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.