Glycerol, sucrose, and other diol-containing reagents are not inert components in in vitro incubations containing aminoacyl-tRNA.
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% (v/v) doubled the rate of deacylation for several aa-tRNAs and peptidyl-tRNAs, including fMet-tRNAMetf, while 1% (v/v) 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 X GTP. In addition, the extent of tRNA aminoacylation was reduced when glycerol was present at concentrations above 2% (v/v). Thus, glycerol and sucrose are not necessarily inert or neutral additions to an in vitro incubation.
D000818:Animals; D055598:Chemical Phenomena; D002621:Chemistry; D004926:Escherichia coli; D005990:Glycerol; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007202:Indicators and Reagents; D012343:RNA, Transfer; D012346:RNA, Transfer, Amino Acyl; D011817:Rabbits; D012441:Saccharomyces cerevisiae; D013395:Sucrose; D013696:Temperature
D007202:Indicators and Reagents; D012346:RNA, Transfer, Amino Acyl; C019037:tRNA, peptidyl-; D013395:Sucrose; D012343:RNA, Transfer; D005990:Glycerol