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. 2007 Jul;13(7):967–973. doi: 10.1261/rna.515707

FIGURE 4.

FIGURE 4.

Characteristics of the C310A mutant protein. (A) Far-UV CD spectra in 0.02 M Tris-HCl buffer (pH 8), 50 mM NaF at a protein concentration of 10 μM or 8 μM for Trm4p (continuous line) and C310A-Trm4p (dashed line), respectively. (B) Comparison of the affinity constants of Trm4p and C310A-Trm4p for yeast tRNAPhe and mini-tRNAPhe using a gel retardation assay with labeled RNA and different concentrations of proteins. (C) SDS-PAGE analysis testing the formation of a covalent complex of C310A-Trm4p with 36-mer FC-mini-RNA substrate analog after Coomassie blue staining. (Lane 1) Molecular weight markers; 3 μg of purified Trm4p (lane 2) alone or (lane 3) in the presence of FC-mini-RNA and SAM; 3 μg of purified C310A-Trm4p (lane 4) alone or (lane 5) in the presence of FC-mini-RNA and SAM.