title: Bisphosphonate mRNA cap analog attached to Sepharose for affinity chromatography of decapping enzymes.
abstract: 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.
authors.affiliation: Division of Biophysics, Institute of Experimental Physics, University of Warsaw, 02-089 Warsaw, Poland.
authors.forename: Sylwia A
authors.initials: SA
authors.lastname: Szczepaniak
authors.forename: Jacek
authors.initials: J
authors.lastname: Jemielity
authors.forename: Joanna
authors.initials: J
authors.lastname: Zuberek
authors.forename: Joanna
authors.initials: J
authors.lastname: Kufel
authors.forename: Edward
authors.initials: E
authors.lastname: Darzynkiewicz
mesh_terms: D000818:Animals; D029681:Arabidopsis Proteins / Q000302:isolation & purification; D002846:Chromatography, Affinity; D002847:Chromatography, Agarose; D004164:Diphosphonates / Q000737:chemistry*; D039561:Eukaryotic Initiation Factor-4E / Q000302:isolation & purification; D051379:Mice; D011755:Pyrophosphatases / Q000302:isolation & purification*; D012314:RNA Cap Analogs / Q000138:chemical synthesis* / Q000737:chemistry; D039381:RNA Cap-Binding Proteins / Q000302:isolation & purification*; D012685:Sepharose / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000737:chemistry
chemical_list: D029681:Arabidopsis Proteins; D004164:Diphosphonates; D039561:Eukaryotic Initiation Factor-4E; D012314:RNA Cap Analogs; D039381:RNA Cap-Binding Proteins; C541622:m7GpCH2pp-sepharose; D012685:Sepharose; D011755:Pyrophosphatases