Figure 4.
Affinity of PUS7 for cytoplasmic tRNAGln. The affinity of different constructs of PUS7 for cy-tRNAGln was determined using nitrocellulose filtration. About 20 nM of cytoplasmic (cy) tRNAGln was incubated in presence of increasing concentration of protein for 10 min in triplicate. The amount of tRNAGln bound to PUS7 in pmol was determined by nitrocellulose filtration and is represented as average with one standard deviation (n = 3−6). The dissociation constants (KD) were determined by fitting the data to a hyperbolic equation (smooth lines) and are summarized in Table 3.