TY - JOUR

T1 - Membrane anchoring facilitates colocalization of enzymes in plant cytochrome P450 redox systems

AU - Laursen, Tomas

AU - Lam, Hiu Yue Monatrice

AU - Sorensen, Kasper Kildegaard

AU - Tian, Pengfei

AU - Hansen, Cecilie Cetti

AU - Groves, Jay T.

AU - Jensen, Knud Jorgen

AU - Christensen, Sune M.

PY - 2021

Y1 - 2021

N2 - Plant metabolism depends on cascade reactions mediated by dynamic enzyme assemblies known as metabolons. In this context, the cytochrome P450 (P450) superfamily catalyze key reactions underpinning the unique diversity of bioactive compounds. In contrast to their soluble bacterial counterparts, eukaryotic P450s are anchored to the endoplasmic reticulum membrane and serve as metabolon nucleation sites. Hence, membrane anchoring appears to play a pivotal role in the evolution of complex biosynthetic pathways. Here, a model membrane assay enabled characterization of membrane anchor dynamics by single molecule microscopy. As a model system, we reconstituted the membrane anchor of cytochrome P450 oxidoreductase (POR), the ubiquitous electron donor to all microsomal P450s. The transmembrane segment in the membrane anchor of POR is relatively conserved, corroborating its functional importance. We observe dynamic colocalization of the POR anchors in our assay suggesting that membrane anchoring might promote intermolecular interactions and in this way impact assembly of metabolic multienzyme complexes.Laursen et al. investigates the role of membrane anchoring for the organization and efficiency of P450-driven plant metabolism. They report an original experimental design to monitor interactions between anchoring segments of membrane-bound enzymes at the single molecule level, using the cytochrome P450 oxidoreductase (POR) transmembrane segment as a model, and propose that membrane anchoring has played a key role in the evolution of P450- based metabolic pathways in plants.

AB - Plant metabolism depends on cascade reactions mediated by dynamic enzyme assemblies known as metabolons. In this context, the cytochrome P450 (P450) superfamily catalyze key reactions underpinning the unique diversity of bioactive compounds. In contrast to their soluble bacterial counterparts, eukaryotic P450s are anchored to the endoplasmic reticulum membrane and serve as metabolon nucleation sites. Hence, membrane anchoring appears to play a pivotal role in the evolution of complex biosynthetic pathways. Here, a model membrane assay enabled characterization of membrane anchor dynamics by single molecule microscopy. As a model system, we reconstituted the membrane anchor of cytochrome P450 oxidoreductase (POR), the ubiquitous electron donor to all microsomal P450s. The transmembrane segment in the membrane anchor of POR is relatively conserved, corroborating its functional importance. We observe dynamic colocalization of the POR anchors in our assay suggesting that membrane anchoring might promote intermolecular interactions and in this way impact assembly of metabolic multienzyme complexes.Laursen et al. investigates the role of membrane anchoring for the organization and efficiency of P450-driven plant metabolism. They report an original experimental design to monitor interactions between anchoring segments of membrane-bound enzymes at the single molecule level, using the cytochrome P450 oxidoreductase (POR) transmembrane segment as a model, and propose that membrane anchoring has played a key role in the evolution of P450- based metabolic pathways in plants.

KW - P450 REDUCTASE

KW - TRACKING REVEALS

KW - SINGLE-MOLECULE

KW - NADPH

KW - PROTEIN

KW - ORGANIZATION

KW - INSERTION

KW - SEQUENCE

KW - OXIDOREDUCTASE

KW - BIOSYNTHESIS

U2 - 10.1038/s42003-021-02604-1

DO - 10.1038/s42003-021-02604-1

M3 - Journal article

C2 - 34504298

VL - 4

JO - Communications Biology

JF - Communications Biology

SN - 2399-3642

M1 - 1057

ER -