Glycerol, sucrose, and other diol-containing reagents are not inert components in in vitro incubations containing aminoacyl-tRNA

Add time:09/07/2019    Source:sciencedirect.com

The addition of glycerol, sucrose, or other diol-containing reagents to solutions of aminoacyl-tRNA (aa-tRNA) substantially increased the rate of hydrolysis of the aminoacyl ester bond. Glycerol at 4.9% (vv) doubled the rate of deacylation ofr several aa-tRNAs and peptidyl-tRNAs, including fMet-tRNAfMet, while 1% (vv) glycerol increased the deacylation rate by 20%. This effect was not caused by a nuclease contamination, and tRNA deacylated in the presence of glycerol could be fully recharged. The deacylation of aa-tRNA was accelerated by glycerol and sucrose even in the presence of EF-Tu·GTP. In addition, the extent of tRNA aminoacylation was reduced when glycerol was present at concentrations above 2% (vv). Thus, glycerol and sucrose are not necessarily inert or neutral additions to an in vitro incubation.

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