Formulation: 50 mM Tris, pH 7.5, with 200 mM sodium chloride and 20% glycerol
Purity: ≥90% estimated by SDS-PAGE
Formula Weight: 0
Shelf life (days): 365
Notes: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is an enveloped positive-stranded RNA virus and the causative agent of COVID-19, a primarily respiratory illness characterized by fever, cough, and shortness of breath that can lead to life-threatening complications.{53462,53459,53461,55257,55258} The SARS-CoV-2 genome contains approximately 30 kilobases and 14 open reading frames (ORFs) that encode four structural proteins: spike, envelope, membrane, and nucleocapsid, as well as 16 non-structural proteins and 9 accessory factors.{57176} SARS-CoV-2 non-structural protein 14 (nsp14), also known as guanine-N7 methyltransferase (guanine-N7 MTase), is encoded within ORF1ab with an amino acid sequence that displays few mutations and is highly conserved across coronaviruses and SARS-CoV-2 variants.{57176,69748,67303} It is a bifunctional enzyme having an N-terminal exoribonuclease domain (ExoN domain) and a C-terminal methyltransferase domain.{57176,69748} SARS-CoV-2 nsp14 forms an exonuclease complex with the cofactor nsp10, which binds to and stabilizes the disordered ExoN domain and is a necessary interaction for accurate exoribonuclease and RNA strand proofreading activities.{67303,57176,69748} It also forms a ternary methylation complex with nsp10 and nsp16 methyltransferase, also known as 2ʹ-O-methyltransferase (2ʹ-O-MTase), and catalyzes guanosine methylation in the viral mRNA 5ʹ-end cap prior to the terminal ribose methylation catalyzed by 2ʹ-O-MTase.{57176,52794} SARS-CoV-2 nsp14 inhibits IFN-β reporter gene activation induced by the retinoic acid-inducible gene I caspase activation and recruitment domain (RIG-I 2CARD domain) in HEK293FT cells, as well as inhibits global protein translation and expression of IFN-stimulated genes (ISGs) in HEK293T cells expressing SARS-CoV-2 nsp14.{57182,69748}
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