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Assay for Rab Geranylgeranyltransferase Using Size Exclusion Chromatography

✍ Scribed by Vo Si Hung; Peter Low; Ewa Swiezewska

Publisher
Elsevier Science
Year
2001
Tongue
English
Weight
124 KB
Volume
289
Category
Article
ISSN
0003-2697

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✦ Synopsis

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.

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