An affinity column for the purification of prenyltransferases.
Farnesyl pyrophosphate synthetase (EC 2.5.1.1) from chicken liver, pig liver, and yeast has been purified to homogeneity in a single chromatographic step by affinity chromatography. The affinity ligand, geranylmethylphosphonophosphate, is linked to Affi-Gel 10 through the phosphonophosphate moiety. The affinity gel is stable chemically and the internal phosphonophosphate linkage is not hydrolyzed by nonspecific phosphatases. A single column has been used repeatedly for over a year with no degradation in its performance. A typical purification only requires 2 days and gives a 500- to 600-fold purification of enzyme from a crude ammonium sulfate precipitate.
D000345:Affinity Labels; D000818:Animals; D002645:Chickens; D002846:Chromatography, Affinity; D004122:Dimethylallyltranstransferase / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel; D008024:Ligands; D008099:Liver / Q000201:enzymology; D012441:Saccharomyces cerevisiae / Q000201:enzymology; D013552:Swine; D014166:Transferases / Q000302:isolation & purification*
D000345:Affinity Labels; D008024:Ligands; D014166:Transferases; D004122:Dimethylallyltranstransferase