Bisphosphonate mRNA cap analog attached to Sepharose for affinity chromatography of decapping enzymes.
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.
Szczepaniak
Sylwia A
SA
Division of Biophysics, Institute of Experimental Physics, University of Warsaw, 02-089 Warsaw, Poland.
Jemielity
Jacek
J
Zuberek
Joanna
J
Kufel
Joanna
J
Darzynkiewicz
Edward
E
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
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