Synthesis and Characterization of Conjugates Formed between Periodate-Oxidized Ribonucleotides and Amine-Containing Fluorophores

Ronald E. Hileman, Kay Martin Parkhurst, Naba K. Gupta, Lawrence J Parkhurst

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

The synthesis and purification of new fluorescently labeled derivatives of GDP and ATP are described. The fluorescent groups are coupled initially through amine-containing linker arms to periodate-oxidized nucleotides. Reduction of the initial product yields primarily a six-membered morpholine-like ring. Fluorescein-labeled GDP, rhodamine-labeled GDP, and fluorescein-labeled ATP were characterized by absorbance spectroscopy and TLC. NMR and FAB-MS studies were carried out on a single nucleotide derivative formed by reacting periodate-oxidized guanosine and benzylamine with subsequent reduction to establish the modification to the ribose moiety. The synthesis of the guanosine—benzylamine conjugate led to a mixture of products that were separated. The predominant product (70%) resulted in conversion of the ribose moiety to a six-membered morpholine-like ring having no hydroxyl group, and the minor product (30%) resulted in an open ring structure having one hydroxyl. When NaCNBH3 was used as the sole reductant, only the product with the morpholine-like ring was formed. These probes were prepared for use in solution studies of the interactions of eukaryotic initiation factor-2 with other components of mammalian protein synthesis initiation.

Original languageEnglish (US)
Pages (from-to)436-444
Number of pages9
JournalBioconjugate Chemistry
Volume5
Issue number5
DOIs
StatePublished - Sep 1 1994

Fingerprint

Ribonucleotides
Fluorophores
Amines
Ribose
Adenosinetriphosphate
Nucleotides
Fluorescein
Hydroxyl Radical
Adenosine Triphosphate
Eukaryotic Initiation Factor-2
Derivatives
Rhodamines
Guanosine
Reducing Agents
Purification
Spectrum Analysis
Nuclear magnetic resonance
Spectroscopy
Proteins
metaperiodate

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Biomedical Engineering
  • Pharmacology
  • Pharmaceutical Science
  • Organic Chemistry

Cite this

Synthesis and Characterization of Conjugates Formed between Periodate-Oxidized Ribonucleotides and Amine-Containing Fluorophores. / Hileman, Ronald E.; Parkhurst, Kay Martin; Gupta, Naba K.; Parkhurst, Lawrence J.

In: Bioconjugate Chemistry, Vol. 5, No. 5, 01.09.1994, p. 436-444.

Research output: Contribution to journalArticle

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