3GSY

Structure of berberine bridge enzyme in complex with dehydroscoulerine

Summary for 3GSY

• Entry DOI: 10.2210/pdb3gsy/pdb
• Related: 3D2D 3D2H 3D2J 3FW7 3FW8 3FW9
• Descriptor: Reticuline oxidase; Berberine bridge-forming enzyme, beta-D-mannopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose, FLAVIN-ADENINE DINUCLEOTIDE, ... (7 entities in total)
• Functional Keywords: complex with dehydroscoulerine, bicovalent flavinylation, n-glycosylation, p-cresol methyl hydroxylase superfamily, alkaloid metabolism, cytoplasmic vesicle, fad, flavoprotein, glycoprotein, oxidoreductase
• Biological source: Eschscholzia californica (California poppy)
• Cellular location: Cytoplasmic vesicle: P30986
• Total number of polymer chains: 1
• Total formula weight: 59817.28

Authors

Winkler, A.,Macheroux, P.,Gruber, K. (deposition date: 2009-03-27, release date: 2009-06-30, Last modification date: 2023-09-06)

Primary citation

Winkler, A.,Puhl, M.,Weber, H.,Kutchan, T.M.,Gruber, K.,Macheroux, P.
Berberine bridge enzyme catalyzes the six electron oxidation of (S)-reticuline to dehydroscoulerine.
Phytochemistry, 70:1092-1097, 2009

PubMed Abstract: Berberine bridge enzyme catalyzes the stereospecific oxidation and carbon-carbon bond formation of (S)-reticuline to (S)-scoulerine. In addition to this type of reactivity the enzyme can further oxidize (S)-scoulerine to the deeply red protoberberine alkaloid dehydroscoulerine albeit with a much lower rate of conversion. In the course of the four electron oxidation, no dihydroprotoberberine species intermediate was detectable suggesting that the second oxidation step leading to aromatization proceeds at a much faster rate. Performing the reaction in the presence of oxygen and under anoxic conditions did not affect the kinetics of the overall reaction suggesting no strict requirement for oxygen in the oxidation of the unstable dihydroprotoberberine intermediate. In addition to the kinetic characterization of this reaction we also present a structure of the enzyme in complex with the fully oxidized product. Combined with information available for the binding modes of (S)-reticuline and (S)-scoulerine a possible mechanism for the additional oxidation is presented. This is compared to previous reports of enzymes ((S)-tetrahydroprotoberberine oxidase and canadine oxidase) showing a similar type of reactivity in different plant species.

PubMed: 19570558
DOI: 10.1016/j.phytochem.2009.06.005

Experimental method

X-RAY DIFFRACTION (1.63 Å)