New affinity resin for purification of cap-binding proteins.
Cap binding proteins, which recognize the cap structure present at 5' termini of RNA polymerase II transcripts, have been routinely isolated and purified using affinity resins with mononucleotide cap analogs attached. Here we present a new methodology in which dinucleotide cap analog, m7GpppG, has been linked to the EAH-Sepharose. The method is based on derivatization of 2',3'-cis diol of the second nucleotide within the cap structure, with levulinic acid, and subsequent coupling of resulted acetal through its carboxylic group with aminohexyl-agarose.
Jankowska-Anyszka
Marzena
M
Department of Chemistry, Warsaw University, 02093 Warsaw, Pasteura 1, Poland. marzena@chem.uw.edu.pl
Nogalski
Maciej
M
Darzynkiewicz
Edward
E
D002352:Carrier Proteins / Q000302:isolation & purification*; D002845:Chromatography / Q000295:instrumentation*; D006868:Hydrolysis; D007475:Ion Exchange Resins / Q000737:chemistry*; D007982:Levulinic Acids / Q000737:chemistry; D008024:Ligands; D008956:Models, Chemical; D017434:Protein Structure, Tertiary; D012315:RNA Caps / Q000737:chemistry*; D012685:Sepharose / Q000737:chemistry*; D013696:Temperature
D002352:Carrier Proteins; D007475:Ion Exchange Resins; D007982:Levulinic Acids; D008024:Ligands; D012315:RNA Caps; D012685:Sepharose; C032246:levulinic acid