Isolation of transfer RNA isoacceptors by chromatography on dihydroxyboryl-substituted cellulose, polyacrylamide, and glass

✍ Scribed by R.E. Duncan; P.T. Gilham

• Publisher: Elsevier Science
• Year: 1975
• Tongue: English
• Weight: 537 KB
• Volume: 66
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(75)90620-x

✦ Synopsis

Polyacrylamide and porous-glass supports containing the dihydroxyborylphenyl group can be prepared by a method similar to that used in the synthesis of N-[ N'-(m-dihydroxyborylphenyl)succinamyl]aminoethylcellulose.

The reaction of aminoethylpolyacrylamide or amino-substituted glass with N-(m-dihydroxyborylphenyl)succinamic acid in the presence of N-cyclohexyl-N'-p-(4-methylmorpholinium) ethylcarbodiimide yields products which, together with the cellulose derivative, are all capable of binding tRNA dissolved in buffers at pH 8.7. The demonstration that bound tRNA can be released with sorbitol solutions or with low pH buffers, together with studies on the binding of tRNA species that contain chemically modified 3'-terminals, indicate that the predominant binding mechanism consists of cyclic complex formation between the immobilized dihydroxyboryl groups and the 3'-terminal cis-dial groups of the tRNA molecules. Aminoacylated tRNA does not bind under the conditions necessary to bind tRNA and this permits the isolation of specific tRNA isoacceptors. The purification of tRNAPhe and the partial purification of tRNAG1" and tRNATrp are described.