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Molecular recognition of amino acids by RNA aptamers: The evolution into an L-tyrosine binder of a dopamine-binding RNA motif

Published online by Cambridge University Press:  01 April 2000

CECILIA MANNIRONI ,
CHIARA SCERCH ,
PAOLO FRUSCOLONI  and
GLAUCO P. TOCCHINI-VALENTINI
CECILIA MANNIRONI
Affiliation:
Istituto di Biologia Cellulare, Consiglio Nazionale delle Ricerche, “Adriano Buzzati-Traverso” Campus, I-00016 Monterotondo Scalo, Rome, Italy
CHIARA SCERCH
Affiliation:
Istituto di Biologia Cellulare, Consiglio Nazionale delle Ricerche, “Adriano Buzzati-Traverso” Campus, I-00016 Monterotondo Scalo, Rome, Italy
PAOLO FRUSCOLONI
Affiliation:
Istituto di Biologia Cellulare, Consiglio Nazionale delle Ricerche, “Adriano Buzzati-Traverso” Campus, I-00016 Monterotondo Scalo, Rome, Italy
GLAUCO P. TOCCHINI-VALENTINI
Affiliation:
Istituto di Biologia Cellulare, Consiglio Nazionale delle Ricerche, “Adriano Buzzati-Traverso” Campus, I-00016 Monterotondo Scalo, Rome, Italy
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Abstract

We report the evolution of an RNA aptamer to change its binding specificity. RNA aptamers that bind the free amino acid tyrosine were in vitro selected from a degenerate pool derived from a previously selected dopamine aptamer. Three independent sequences bind tyrosine in solution, the winner of the selection binding with a dissociation constant of 35 μM. Competitive affinity chromatography with tyrosine-related ligands indicated that the selected aptamers are highly L-stereo selective and also recognize L-tryptophan and L-dopa with similar affinity. The binding site was localized by sequence comparison, analysis of minimal boundaries, and structural probing upon ligand binding. Tyrosine-binding sites are characterized by the presence of both tyrosine (UAU and UAC) and termination (UAG and UAA) triplets.

Keywords

affinity chromatographyamino acidRNA aptamersselection-amplification
Type
Research Article
Information
RNA , Volume 6 , Issue 4 , April 2000 , pp. 520 - 527
Copyright
© 2000 RNA Society

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