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PMID: 10786843 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

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

RNA (New York, N.Y.) ·Vol. 6 ·No. 4 ·2000-04-00 ·Pages 520-7

Mannironi C, Scerch C, Fruscoloni P, Tocchini-Valentini GP

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 microM. 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.

MeSH Terms

Base Sequence Binding Sites Chromatography, Affinity Cloning, Molecular Directed Molecular Evolution Dopamine/chemistry,metabolism Ligands Molecular Probe Techniques Nucleic Acid Conformation Oligoribonucleotides/chemistry,genetics,metabolism Phenylalanine/genetics,metabolism RNA/chemistry,genetics,metabolism Substrate Specificity Thermodynamics Tryptophan/genetics,metabolism Tyrosine/genetics,metabolism

Chemicals

Ligands Oligoribonucleotides Tyrosine Phenylalanine RNA Tryptophan Dopamine

Authors & Affiliations

4 authors, click to expand affiliations / ORCID

Mannironi C

Istituto di Biologia Cellulare, Consiglio Nazionale delle Ricerche, Rome, Italy.

Scerch C

Fruscoloni P

Tocchini-Valentini G P

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16 references, click to expand

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Article Info

Journal

RNA (New York, N.Y.)

Abbr.

RNA

ISSN

1355-8382

DOI

10.1017/s1355838200991763

Published

2000-04-00

Pages

520-7

Language

English

Region

United States

NLM ID

9509184

PMCID

PMC1369933

Subset

External Links

PubMed source DOI: 10.1017/s1355838200991763 PMC Full Text View journal details

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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.

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