Geranylgeranyl pyrophosphate ammonium salt solution

Описание

Geranylgeranyl pyrophosphate (GGPP) is a potent endogenous regulator of Hmg2p degradation.This work was launched by the unexpected observation that GGPP addition directly to living yeast cultures caused high potency and specific stimulation of Hmg2p degradation. This effect of GGPP was not recapitulated by FPP,GGOH, or related isoprenoids. GGPP-caused Hmg2p degradation met all the criteria for the previously characterized endogenous signal.The action of added GGPP did not require production of endogenous sterol molecules, indicating that it did not act by causing the build-up of an endogenouspathway signal.

Использование

Geranylgeranyl pyrophosphate ammonium salt has been used:
in the preparation of 2.5X reaction buffer for Rab GGTase assay
to add to the culture to further drive Hmg2 degradation
as a substrate to carryout in vitro enzymatic assays for the characterization of the prenyltransferases
to reverse the action of lovastatin and also used to examine the potential role of GGPP in the study to assess the effects of exposing the melanoma cell lines in the angiogenesis model as a co-culture to lovastatin?in vitro

Биохимия/физиол Действия

Geranylgeranyl pyrophosphate (GGPP) is a major enzyme that participates in the secondary metabolism of chloroplast. GGPP synthase plays a key role in the biosynthesis of terpenoid. Protein geranylgeranylation, mediated by geranylgeranyl pyrophosphate participates in the development of the ventricular chamber. It also serves as a stage-specific signal to modulate the formation of cardiac cytoarchitecture the time of mid-gestation.

Методы очистки

Purify the pyrophosphate by countercurrent distribution between two phases of a butanol/isopropyl ether/ammonia/water mixture (15:5:1:19) (v/v), or by chromatography on DEAE-cellulose (linear gradient of 0.02M KCl in 1mM Tris buffer, pH 8.9). Alternatively purify it through a column of Dowex 1-x8 (formate form previously washed with MeOH) and eluted with a linear gradient of 0.053—0.43M ammonium formate in a total volume of 300mL of MeOH. The purity can be checked by TLC on Silica gel G on buffered plates (pH 6.5), eluted with CHCl3/MeOH/H2O (60:40:9) and developed with I2 vapour. Store it as a powder at 0o. [Altman et al. J Am Chem Soc 94 3257 1972.] Itis more stable as the di(tri-n-butylammonium)hydrogen phosphate salt which can be obtained from the acid by evaporation in a rotary evaporator below 32o [Upper & West J Biol Chem 242 3285 1967]. [Gregonis & Rilling Biochemistry 13 1538 1974, Gregonis & Rilling Biochem Biophys Res Commun 54 449 1973.]