Abstract
To broaden the scope of existing methods based on 19F nucleotide labeling, we developed a new method for the synthesis of fluorophosphate (oligo)nucleotide analogs containing an O to F substitution at the terminal position of the (oligo)phosphate moiety and evaluated them as tools for 19F NMR studies. Using three efficient and comprehensive synthetic approaches based on phosphorimidazolide chemistry and tetra-n-butylammonium fluoride, fluoromonophosphate or fluorophosphate imidazolide as fluorine sources, we prepared over 30 fluorophosphate-containing nucleotides, varying in nucleobase type (A, G, C, U, m7G), phosphate chain length (from mono- to tetra-) and presence of additional phosphate modifications (thio-, borano-, imido-, methylene-). Using fluorophosphate imidazolide as fluorophosphorylating reagent for 5'-phosphorylated oligos we also synthesized oligonucleotide 5'-(2-fluorodiphosphates), which are potentially useful as 19F NMR hybridization probes. The compounds were characterized by 19F NMR, and evaluated as 19F NMR molecular probes. We found that fluorophosphate nucleotide analogs can be used to monitor activity of enzymes with various specificities and metal-ion requirements, including human DcpS enzyme - a therapeutic target for spinal muscular atrophy. The compounds can also serve as reporter ligands for protein binding studies, as exemplified by studying interaction of fluorophosphate mRNA cap analogs with eukaryotic translation initiation factor (eIF4E).
PMID: 25816092 [PubMed - as supplied by publisher]
[NMR paper] Toward a Biorelevant Structure of Protein Kinase C Bound Modulators: Design, Synthesis, and Evaluation of Labeled Bryostatin Analogs for Analysis with REDOR NMR Spectroscopy.
Toward a Biorelevant Structure of Protein Kinase C Bound Modulators: Design, Synthesis, and Evaluation of Labeled Bryostatin Analogs for Analysis with REDOR NMR Spectroscopy.
Toward a Biorelevant Structure of Protein Kinase C Bound Modulators: Design, Synthesis, and Evaluation of Labeled Bryostatin Analogs for Analysis with REDOR NMR Spectroscopy.
J Am Chem Soc. 2015 Feb 24;
Authors: Loy BA, Lesser AB, Staveness D, Billingsley KL, Cegelski L, Wender PA
Abstract
Protein kinase C (PKC) modulators are currently of great...
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02-25-2015 05:56 PM
Characterization of Sarcoplasmic Reticulum Ca(2+) ATPase Nucleotide Binding Domain Mutants using NMR spectroscopy.
Characterization of Sarcoplasmic Reticulum Ca(2+) ATPase Nucleotide Binding Domain Mutants using NMR spectroscopy.
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Biochem Biophys Res Commun. 2010 Dec 24;
Authors: Myint W, Gong Q, Ahn J, Ishima R
Sarcoplasmic reticulum Ca(2+) ATPase (SERCA) is essential for muscle function by transporting Ca(2+) from the cytosol into the sarcoplasmic reticulum through ATP hydrolysis. In this report, the effects of substitution mutations on the...
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12-29-2010 04:04 PM
[NMR paper] Biochemical characterization and NMR studies of the nucleotide-binding domain 1 of mu
Biochemical characterization and NMR studies of the nucleotide-binding domain 1 of multidrug-resistance-associated protein 1: evidence for interaction between ATP and Trp653.
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Biochem J. 2003 Dec 15;376(Pt 3):749-56
Authors: Ramaen O, Masscheleyn S, Duffieux F, Pamlard O, Oberkampf M, Lallemand JY, Stoven V, Jacquet E
Multidrug-resistance-associated...
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11-24-2010 09:16 PM
[NMR paper] NMR studies on the structure/function correlations of T-cell-epitope analogs from per
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Eur J Biochem. 1998 Jun 1;254(2):313-7
Authors: Scarselli M, Esposito G, De Magistris MT, Domenighini M, Rappuoli R, Burroni G, Bernini A, Niccolai N
A synthetic tridecapeptide, corresponding to the 30-42 fragment of the S1 subunit of pertussis toxin, has been structurally characterised by using NMR spectroscopy. The molecule...
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11-17-2010 11:06 PM
Oligo design tools for gene synthesis
Oligo design tools for gene synthesis
1 DNAWorks: http://helixweb.nih.gov/dnaworks/
2 TmPrime: http://prime.ibn.a-star.edu.sg/
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08-25-2010 04:37 AM
[NMR paper] NMR studies of fluororetinol analogs complexed to two homologous rat cellular retinol
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Biochim Biophys Acta. 1994 Sep 21;1208(1):136-44
Authors: Rong D, Lovey AJ, Rosenberger M, d'Avignon DA, Li E
Comparative 19F-NMR studies of fluororetinol analogs with rat cellular retinol binding protein II (CRBP II) and rat cellular retinol-binding protein (CRBP) were performed to probe differences in the binding...
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08-22-2010 03:29 AM
[NMR paper] NMR studies of fluorinated visual pigment analogs.
NMR studies of fluorinated visual pigment analogs.
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Biochem Biophys Res Commun. 1991 Sep 30;179(3):1337-43
Authors: Colmenares LU, Asato AE, Denny M, Mead D, Zingoni JP, Liu RS
The 19F-nmr chemical shift data of isomeric pigments (11-cis and 9-cis) of four vinyl fluororhodopsins and two trifluororhodopsins have been recorded. When compared with model protonated Schiff...
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08-21-2010 11:12 PM
[NMR paper] NMR studies of fluorinated visual pigment analogs.
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http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--linkinghub.elsevier.com-ihub-images-PubMedLink.gif Related Articles NMR studies of fluorinated visual pigment analogs.
Biochem Biophys Res Commun. 1991 Sep 30;179(3):1337-43
Authors: Colmenares LU, Asato AE, Denny M, Mead D, Zingoni JP, Liu RS
The 19F-nmr chemical shift data of isomeric pigments (11-cis and 9-cis) of four vinyl fluororhodopsins and two trifluororhodopsins have been recorded. When compared with model protonated Schiff...
Synthesis of Fluorophosphate Nucleotide Analogs and Their Characterization as Tools for 19F NMR Studies.
Linear Mode
