An in vivo-in vitro alkaline DNA unwinding assay for hepatic DNA damage: comparison with the alkaline sucrose gradient centrifugation technique |
| |
Authors: | R D Storer R B Conolly |
| |
Affiliation: | Department of Chemistry, University of Oklahoma, Norman, Oklahoma 73019 USA |
| |
Abstract: | 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. |
| |
Keywords: | glycerol sucrose aminoacyl-tRNA aminoacylation elongation factor EF-Tu elongation factor eEF-Tu |
本文献已被 ScienceDirect PubMed 等数据库收录! |
|