Efficient cleavage of problematic tobacco etch virus (TEV)-protein arginine methyltransferase constructs.
Protein arginine methyltransferases (PRMTs) are enzymes that are involved in many biological processes. Several studies have shown that the identity of the N-terminal fusion tag affects the substrate selectivity of PRMTs. Therefore, to accurately study substrate recognition, it is imperative that a tagless PRMT be used. However, cleavage of tagged PRMTs has been problematic. We have developed a successful method by which untagged PRMTs can be made using a tobacco etch virus (TEV) cleavage site at the N-terminal domain. This method may be useful for cleaving other challenging target proteins that have the TEV protease recognition site.
Suh-Lailam
Brenda B
BB
Department of Chemistry and Biochemistry, Utah State University, Logan, UT 84322, USA.
Hevel
Joan M
JM
D001665:Binding Sites / Q000235:genetics; D010450:Endopeptidases / Q000737:chemistry; D017800:Potyvirus / Q000201:enzymology* / Q000235:genetics; D011484:Protein-Arginine N-Methyltransferases / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000737:chemistry
D011993:Recombinant Fusion Proteins; D011484:Protein-Arginine N-Methyltransferases; D010450:Endopeptidases; C099488:TEV protease