Direct synthesis of aminoacyl-tRNA occurs using α-N-tBoc-protected aminoacyl phosphates and lanthanum salts. Deprotection of aminoacyl tRNA is essential prior to translation; however, the conditions of tBoc deprotection causes tRNA degradation. It was found that α-N-pentenoyl-protected aminoacyl phosphates, deprotected under mild conditions, are effectively used in lanthanum-mediated acylation of tRNA analogs. This provides an alternative route for aminoacyl-tRNA synthesis that maintains tRNA structure. Also, it was determined that α-N-deprotection of aminoacyl phosphates prior to aminoacylation still produces aminoacylated tRNA analogs. This establishes that acyl phosphates activate amino acids without inducing self-condensation, presumably due to electrostatic repulsion. Direct quantification of lanthanum-mediated tRNA aminoacylation was additionally undertaken utilizing a radiolabelled tRNA assay. From this, it was shown that lanthanum-mediated acylation does not promote deacylation and degradation of tRNA. These results have provided insight into lanthanum-mediated acylation of tRNA, ultimately allowing for use of the reagent in ribosomal translation.
| Identifer | oai:union.ndltd.org:TORONTO/oai:tspace.library.utoronto.ca:1807/18143 |
| Date | 15 December 2009 |
| Creators | Andrusiak, Tara |
| Contributors | Kluger, Ronald |
| Source Sets | University of Toronto |
| Language | en_ca |
| Detected Language | English |
| Type | Thesis |
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