Boronate affinity adsorption of RNA: Possible role of conformational changes

Narinderjeet Singh, Richard C. Willson

Research output: Contribution to journalArticle

33 Scopus citations

Abstract

Batch equilibrium adsorption isotherm determination is used to characterize the adsorption of mixed yeast RNA on agarose-immobilized m-aminophenylboronic acid. It is shown that the affinity-enhancing influence of divalent cations depends strongly on the precise nature of the cation used, with barium being far more effective than the conventionally-used magnesium. This adsorption-promoting influence of barium is suggested to arise primarily from ionic influences on the structure and rigidity of the RNA molecule, as the adsorption of ribose-based small molecules is not similarly affected. The substitution of barium for the standard magnesium counterion does not greatly promote the adsorption of DNA, implying that the effect is specific to RNA and may be useful in boronate-based RNA separations. RNA adsorption isotherms exhibit sharp transitions as functions of temperature, and these transitions occur at different temperatures with Mg2+ and with Ba2+. Adsorption affinity and capacity were found to increase markedly at lower temperatures, suggestive of an enthalpically favored interaction process. The stoichiometric displacement parameter, Z, in Ba2+ buffer is three times the value in Mg2+ buffer, and is close to unity. Copyright (C) 1999 Published by Elsevier Science B.V.

Original languageEnglish (US)
Pages (from-to)205-213
Number of pages9
JournalJournal of Chromatography A
Volume840
Issue number2
DOIs
StatePublished - Apr 30 1999

Keywords

  • Adsorption isotherms
  • Inorganic cations
  • m-Aminophenylboronic acid
  • RNA

ASJC Scopus subject areas

  • Analytical Chemistry

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