Chapter 7
Transportomics for the Characterization of Plant
Apocarotenoid Transmembrane Transporters
Olivia Costantina Demurtas, Rita de Brito Francisco, Enrico Martinoia,
and Giovanni Giuliano
Abstract
Apocarotenoids are carotenoid derivatives produced by the nonenzymatic or enzymatic cleavage of carotenoids, followed by different enzymatic modifications. In plants, apocarotenoids play different roles, such
as attraction of pollinators and seeds dispersal, defense against pathogens and herbivores, protection against
photo-oxidative stresses, stimulation and inhibition of plant growth and regulation of biological processes
in the case of phytohormones abscisic acid and strigolactones. While carotenoids are in general plastidlocalized metabolites, apocarotenoids can reach different final destinations inside or outside the cell. The
mechanisms of apocarotenoid transport through biological membranes have been poorly studied. This
chapter describes a method to characterize transmembrane transporters involved in the transport of polar
and amphipathic apocarotenoids. This protocol was successfully used to in vitro characterize the transport
activity of ATP-binding cassette (ABC) and multidrug and toxic extrusion (MATE) in microsomes isolated
from Saccharomyces cerevisiae expressing these plant transporters.
Key words Carotenoids, Apocarotenoids, ATP-binding cassette (ABC) transporters, Multidrug and
toxic compound extrusion (MATE) transporters, Yeast microsomes, LC-MS
1 Introduction
Apocarotenoids are produced from carotenoids through both
enzymatic and nonenzymatic cleavage. In plants, they play important biological roles as pigments, volatiles and signaling molecules
[1–3]. In mammals, apocarotenoids play roles in signaling and
regulation of metabolism [4].
While the key enzymatic steps that lead to the formation of
apocarotenoids in different plant species have been studied, the
proteins mediating their intra- and intercellular transport are still
poorly investigated. Many apocarotenoids, and their glycosylated
Manuel Rodrı ´guez-Concepcio ´ n and Ralf Welsch (eds.), Plant and Food Carotenoids: Methods and Protocols,
Methods in Molecular Biology, vol. 2083, https://doi.org/10.1007/978-1-4939-9952-1_7,
© Springer Science+Business Media, LLC, part of Springer Nature 2020
Olivia Costantina Demurtas and Rita de Brito Francisco contributed equally to this work.
89
Transportomics for the Characterization of Plant
Apocarotenoid Transmembrane Transporters
Olivia Costantina Demurtas, Rita de Brito Francisco, Enrico Martinoia,
and Giovanni Giuliano
Abstract
Apocarotenoids are carotenoid derivatives produced by the nonenzymatic or enzymatic cleavage of carotenoids, followed by different enzymatic modifications. In plants, apocarotenoids play different roles, such
as attraction of pollinators and seeds dispersal, defense against pathogens and herbivores, protection against
photo-oxidative stresses, stimulation and inhibition of plant growth and regulation of biological processes
in the case of phytohormones abscisic acid and strigolactones. While carotenoids are in general plastidlocalized metabolites, apocarotenoids can reach different final destinations inside or outside the cell. The
mechanisms of apocarotenoid transport through biological membranes have been poorly studied. This
chapter describes a method to characterize transmembrane transporters involved in the transport of polar
and amphipathic apocarotenoids. This protocol was successfully used to in vitro characterize the transport
activity of ATP-binding cassette (ABC) and multidrug and toxic extrusion (MATE) in microsomes isolated
from Saccharomyces cerevisiae expressing these plant transporters.
Key words Carotenoids, Apocarotenoids, ATP-binding cassette (ABC) transporters, Multidrug and
toxic compound extrusion (MATE) transporters, Yeast microsomes, LC-MS
1 Introduction
Apocarotenoids are produced from carotenoids through both
enzymatic and nonenzymatic cleavage. In plants, they play important biological roles as pigments, volatiles and signaling molecules
[1–3]. In mammals, apocarotenoids play roles in signaling and
regulation of metabolism [4].
While the key enzymatic steps that lead to the formation of
apocarotenoids in different plant species have been studied, the
proteins mediating their intra- and intercellular transport are still
poorly investigated. Many apocarotenoids, and their glycosylated
Manuel Rodrı ´guez-Concepcio ´ n and Ralf Welsch (eds.), Plant and Food Carotenoids: Methods and Protocols,
Methods in Molecular Biology, vol. 2083, https://doi.org/10.1007/978-1-4939-9952-1_7,
© Springer Science+Business Media, LLC, part of Springer Nature 2020
Olivia Costantina Demurtas and Rita de Brito Francisco contributed equally to this work.
89