TY - JOUR
T1 - How to prove the existence of metabolons?
AU - Bassard, Jean-Étienne André
AU - Halkier, Barbara Ann
PY - 2018/4/1
Y1 - 2018/4/1
N2 - 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.
AB - 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.
KW - Fluorescence correlation spectroscopy
KW - Fluorescence lifetime imaging microscopy
KW - Fluorescence-based protein–protein interaction
KW - Yeast-2-hybrid screen
U2 - 10.1007/s11101-017-9509-1
DO - 10.1007/s11101-017-9509-1
M3 - Journal article
C2 - 29755303
SN - 1568-7767
VL - 17
SP - 211
EP - 227
JO - Phytochemistry Reviews
JF - Phytochemistry Reviews
IS - 2
ER -