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Crystallization and preliminary X‐ray crystallographic analysis of a nonstructural protein 15 mutant from Human coronavirus 229E
Author(s) -
Huo Tong,
Liu Xiang
Publication year - 2015
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 37
ISSN - 2053-230X
DOI - 10.1107/s2053230x15007359
Subject(s) - endoribonuclease , mutant , coronavirus , chemistry , microbiology and biotechnology , open reading frame , virology , crystallography , mutant protein , gene , biology , rna , biochemistry , peptide sequence , medicine , covid-19 , disease , pathology , infectious disease (medical specialty) , rnase p
Nonstructural protein 15 (nsp15), also called endoribonuclease, is a gene product of open reading frame 1b (ORF 1b) in coronaviruses. It is an important enzyme in the transcription/replication process involved in discontinuous negative‐strand RNA synthesis. In this work, mutants of nsp15 from Human coronavirus 229E (HCoV‐229E) were made based on structural analysis of the homologous nsp15s in Severe acute respiratory syndrome coronavirus (SARS‐CoV) and Mouse hepatitis virus (MHV). The I26A/N52A mutant of nsp15 was overexpressed, purified and crystallized, and this mutant led to a trimeric form rather than hexamers or monomers. Crystals of trimeric nsp15 were obtained by the hanging‐drop vapour‐diffusion method using polyethylene glycol as a precipitant and diffracted to 2.5 Å resolution. The crystals belonged to space group C 222 1 , with unit‐cell parameters a = 85.9, b = 137.5, c = 423.1 Å, α = β = γ = 90°.