How to prove the existence of metabolons?

Phytochemistry Reviews : Proceedings of the Phytochemical Society of Europe
Jean-Etienne Bassard, Barbara Ann Halkier

Abstract

Sequential enzymes in biosynthetic pathways are organized in metabolons. It is challenging to provide experimental evidence for the existence of metabolons as biosynthetic pathways are composed of highly dynamic protein-protein interactions. Many different methods are being applied, each with strengths and weaknesses. We will present and evaluate several techniques that have been applied in providing evidence for the orchestration of the biosynthetic pathways of cyanogenic glucosides and glucosinolates in metabolons. These evolutionarily related pathways have ER-localized cytochromes P450 that are proposed to function as anchoring site for assembly of the enzymes into metabolons. Additionally, we have included commonly used techniques, even though they have not been used (yet) on these two pathways. In the review, special attention will be given to less-exploited fluorescence-based methods such as FCS and FLIM. Ultimately, understanding the orchestration of biosynthetic pathways may contribute to successful engineering in heterologous hosts.

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Oct 17, 2018·Natural Product Reports·Camilla KnudsenTomas Laursen
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Methods Mentioned

BETA
co-immunoprecipitation
tandem
fluorescence correlation spectroscopy
Y2H
FCS
Co-IP
transgenic
affinity purification
tandem affinity purification
pull-down

Software Mentioned

FCS
Co
FLIM
IP

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