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Monoclonal Antibody Services> | Flag-tagged proteins were detected using alkaline phosphatase-linked anti-Flag M2 monoclonal antibody (Sigma) and StrepII-tagged proteins were detected using unconjungated rabbit anti-StrepII polyclonal antibody (Genscript USA, Piscataway, NJ) for subsequent alkaline phosphatase-linked goat anti-rabbit IgG (H+L) antibody detection (SouthernBiotech, Birmingham, AL). | Get A Quote |
While cytoplasmic tRNA 2-thiolation protein 1 (Tuc1/Ncs6) and ubiquitin-related modifier-1 (Urm1) are important in the 2-thiolation of 5-methoxycarbonylmethyl-2-thiouridine (mcm5s2U) at wobble uridines of tRNAs in eukaryotes, the biocatalytic roles and properties of Ncs6/Tuc1 and its homologs are poorly understood. Here we present the first report of an Ncs6 homolog of archaea (NcsA of Haloferax volcanii) that is essential for maintaining cellular pools of thiolated tRNA(Lys)UUU and for growth at high temperature. When purified from Hfx. volcanii, NcsA was found to be modified at Lys204 by isopeptide linkage to polymeric chains of the ubiquitin-fold protein SAMP2. The ubiquitin-activating E1 enzyme homolog ... More