US EPA

8175526 

Journal Article 

Protein-independent interaction of divalent metal ions with geranylgeranyl diphosphate and other prenyl diphosphates 

Reuter, B; Knoss, W; Hemmerlin, A; Bach, TJ 

1997 

7 

4 

141-144 

English 

[14C]Geranylgeranyl diphosphate was produced in high yields by a prenyl transferase system extracted from the mutant strain SG4 of Gibberella fujikuroi. When the incubation mixture was subjected to dialysis or ultrafiltration, the radiolabelled product was retained with the high molecular weight fraction. A purification procedure was developed based on acetone precipitation of [14C]geranylgeranyl diphosphate, resolubilisation and subsequent solid phase extraction using a silica gel stationary phase. Controls with solutions of geranylgeranyl diphosphate in buffer demonstrated that the apparent precipitation is not due to an interaction with proteins, but depends on interactions with Mn2+, Mg2+ or other divalent metal ions. Analogous interactions were shown for other prenyl diphosphates and must be considered in design and evaluation of all methods using those substrates.