Assay for Rab geranylgeranyltransferase using size exclusion chromatography.
A simple chromatographic assay for Rab geranylgeranyltransferase (Rab GGTase) has been developed. The method involves separation of the reaction mixture on a Sephadex G-25 superfine minicolumn. Addition of 2-propanol to the assay results in substantial (approximately 90%) decline of formation of noncovalent lipid-protein complexes, increasing reproducibility and reliability of the method. The activity of Rab prenyltransferase was measured in crude and partially purified enzyme preparations from wheat seedlings; measurements for several other plants and rat brain cytosol fractions are also presented. This method can be routinely applied to evaluate the activity of different protein prenyltransferases.
Hung
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VS
Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawinskiego 5a, Warsaw, 02-106, Poland.
Low
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P
Swiezewska
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E
D019883:Alkyl and Aryl Transferases / Q000378:metabolism*; D002850:Chromatography, Gel / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D011994:Recombinant Proteins / Q000378:metabolism
D011994:Recombinant Proteins; D019883:Alkyl and Aryl Transferases; C075607:Rab geranylgeranyltransferase