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Alteration of conserved residues in the SAM-binding motif affects SAM-dependence of methylation by ScTrm10. Methylation activity of each ScTrm10 enzyme with G9-labeled tRNAGly was quantified as described in methods and plots of percent conversion to m1G9 versus [SAM] were fit to Equation (1) to determine K1/2 (SAM) for each enzyme: G206A/G207A (open diamonds; K1/2 = 77.6 ± 9.9 μM), D210K (squares; K1/2 = 91.8 ± 8.7 μM), D210A (circles; K1/2 = 4.3 ± 0.4 μM).
