JBC, Vol. 252, Issue 8, 2560-2565, Apr, 1977
Isolation and characterization of valine transfer RNA from Saccharomyces cerevisiae
S. Aoyagi, O. S. Bhanot and R. W. Chambers
Two procedures for isolating valine tRNA from commercial bakers' yeast were
investigated. The first involved: (a) counter double current distribution;
(b) chromatography on benzoyl-DEAE-cellulose; (c) reverse phase
chromatography on Chromosorb G saturated with trioctylpropylammonium
bromide (Oakridge System 3). The material isolated lacked the 3'-terminal
adenylic acid residue. The second procedure involved the first two steps
above followed by: (a) enzymatic aminoacylation with a partially purified
yeast extract; (b) derivatization with N-phenoxyacetoxysuccinimide; (c)
chromatography on benzoyl-DEAE-cellulose; (d) reverse phase chromatography,
System 3. The product was intact tRNA. It was a mixture of isoacceptors
(59:41) differing by a modification (uracil leads to dihydrouracil) at
position 48. It was free of denatured material; specific activity 1,825
pmol of valine/A260 unit of tRNA. Sequence analysis confirmed the recently
corrected structure (Bonnet, J., Ebel, J. P., Dirheimer, G., Shershneva, L.
P., Krutilina, A. I., Venkstern, T. V., and Bayev, A. A. (1974) Biochimie
56, 1211-1213). A preliminary study of the alkaline hydrolysis of the
7-methylguanosine residue that occurs at position 47 showed that at least
two products are formed instead of only one as usually quoted in the
literature. A rapid, ultramicro, chromatographic system for separating
these products and measuring them quantitatively is described.
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
