A simple in vitro assay to measure the activity of geranylgeranyl diphosphate synthase and other short-chain prenyltransferases
Barja, Maria Victoria (Centre de Recerca en Agrigenòmica)
Rodríguez Concepción, Manuel (Centre de Recerca en Agrigenòmica)

Imprint:	New York : Humana, 2020
Description:	12 pàg.
Abstract:	Most carotenoids are C40 metabolites produced from C20 geranylgeranyl diphosphate (GGPP). The enzymes that produce this precursor, GGPP synthases (GGPPS), are members of the short-chain prenyltransferase (SC-PT) family. SC-PTs are enzymes that catalyze the sequential head-to-tail addition of one or more C5 molecules of isopentenyl diphosphate (IPP) to dimethylallyl diphosphate (DMAPP) with the concomitant release of pyrophosphate (PPi). SC-PTs produce linear isoprenyl diphosphates of up to C20 (GGPP) that serve as precursors for many groups of isoprenoids with a wide range of essential biological functions in Eucarya, Bacteria, and Archaea. Enzymatic analysis of SC-PT activity normally requires complex, laborious and expensive methods such as radioactivity-based assays or liquid chromatography-mass spectrometry (LC-MS). Here we describe a fast and inexpensive spectrophotometric protocol for determining the kinetic parameters of SC-PTs in purified enzyme preparations, using an adapted assay for PPi quantification. We developed the method using the Arabidopsis thaliana GGPPS11 enzyme, which produces geranylgeranyl diphosphate for the synthesis of carotenoids in the chloroplast.
Rights:	Tots els drets reservats.
Language:	Anglès
Series:	Methods in molecular biology ; 2083
Document:	Capítol de llibre ; Versió acceptada per publicar
Subject:	Short-chain prenyltransferases ; Isoprenyl diphosphate synthases ; Enzymatic activity ; Kinetic parameters ; EnzChek kit ; Geranylgeranyl diphosphate synthase ; GGPP ; Isoprenoids ; Carotenoids
Published in:	Plant and Food Carotenoids, 2020, p. 27-38, ISBN 978-1-4939-9951-4

DOI: 10.1007/978-1-4939-9952-1_2

Postprint 16 p, 489.0 KB

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Research literature > UAB research groups literature > Research Centres and Groups (research output) > Experimental sciences > CRAG (Centre for Research in Agricultural Genomics)
Books and collections > Book chapters