Recently, a mammalian tRNA which was previously designated as an opal suppressor seryl-tRNA and phosphoseryl-tRNA was shown to be a selenocysteyl-tRNA (B. J. Lee, P. J. Worland, J. N. Davis, T. C. Stadtman, and D. Hatfield, J. Biol. Chem. 264:9724-9727, 1989). Hence, this tRNA is now designated as selenocysteyl-tRNA[ISrlsJ, and its function is twofold, to serve as (i) a carrier molecule upon which selenocysteine is biosynthesized and (ii) as a donor of selenocysteine, which is the 21st naturally occurring amino acid of protein, to the nascent polypeptide chain in response to specffic UGA codons. In the present study, the selenocysteine tRNA gene was sequenced from Xenopus laevis, Drosophila melanogaster, and Caenorhabditis elegans. The tRNA product of this gene was also identified within the seryl-tRNA population of a number of higher and lower animals, and the human tRNA FSerlsJ gene was used as a probe to identify homologous sequences within genomic DNAs of organisms throughout the animal kingdom. The studies showed that the tRNA[SerJSec gene has undergone evolutionary change and that it is ubiquitous in the animal kingdom.Further, we conclude that selenocysteine-containing proteins, as well as the use of UGA as a codon for selenocysteine, are far more widespread in nature than previously thought.
A microcantilever-based biosensor is described. The enzyme glucose oxidase was immobilized on a micromachined silicon cantilever containing a gold coating, such as those used for atomic force microscopy. Specific, quantifiable deflection of the derivatized cantilevers was observed in the presence of the appropriate analyte. An analysis of the reaction energetics and the expected thermal response of the cantilever indicates that cantilever deflection is not simply a result of reaction-generated heat. This deflection appears to result from surface induced stresses. The combination of a highly specific enzyme and the microcantilever platform provides a unique approach for quantifying enzyme substrates without the complication of sample labeling.
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