Bisphosphonate mRNA cap analog attached to Sepharose for affinity chromatography of decapping enzymes | Zendy

S Szczepaniak | Zendy; Jacek Jemielity | Zendy; Joanna Zuberek | Zendy; Joanna Kufel | Zendy; E. Darzynkiewicz | Zendy

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Bisphosphonate mRNA cap analog attached to Sepharose for affinity chromatography of decapping enzymes

Author(s) -

S Szczepaniak,

Jacek Jemielity,

Joanna Zuberek,

Joanna Kufel,

E. Darzynkiewicz

Publication year - 2008

Publication title -

nucleic acids symposium series

Language(s) - English

Resource type - Journals

eISSN - 1746-8272

pISSN - 0261-3166

DOI - 10.1093/nass/nrn149

Subject(s) - affinity chromatography , sepharose , enzyme , chemistry , messenger rna , chromatography , biochemistry , gene

m(7)GTP-Sepharose is routinely used for cap binding protein isolation. Here we present the synthesis of a new affinity resin containing a mononucleotide cap analog resistant to hydrolysis by DcpS. The resin has been designed in order to identify and purify Arabidopsis thaliana DcpS and other pyrophosphatases. The binding efficiency of the new resin to eIF4E protein was compared with standard m(7)GTP-Sepharose. The utility of non-hydrolysable resin was demonstrated on yeast extract.

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