Development of an Affinity‐Based Proteomic Strategy for the Elucidation of Proanthocyanidin Biosynthesis

Abstract

Affinity and beyond! An affinity chromatography tool based on specific flavanol–protein interactions has been developed to allow the selective purification and identification by proteomic analysis of functional proteins involved in the last steps of proanthocyanidin biosynthesis.

Real‐Time Analysis of Polyphenol–Protein Interactions by Surface Plasmon Resonance Using Surface‐Bound Polyphenols

“…All three interactions are again globally characterized by rather rapid associations and slow dissociations. These last results further confirm that this SPR technique can be used to unveil structure–protein binding relationships with our polyphenolic probes, since the interaction of the LDOX flavonoid enzyme with the catechin probe 3 e was expectedly detected, [18, 28] whereas the same probe gave very weak or no SPR binding signals with Top2α, BSA, collagen, myoglobin, and streptavidin (see Figures 3 and 4, as well as the Supporting Information, Figures S1 and S2).…”

“…For the derivatization of catechin ( 3 ) and epicatechin ( 4 ) through the A ring, the greater inherent nucleophilic character of their C8 centers [17] was exploited to install a propanoic acid tether, by following a five‐step reaction sequence that we previously described [18] . The resulting carboxylic acids 3 f and 4 a [18] were initially intended to be converted to thioester derivatives by using octane‐1,8‐dithiol under standard Steglich‐type esterification conditions in the presence of 1‐ethyl‐3‐(3‐dimethylaminopropyl) carbodiimide (EDCI). However, a δ‐lactonization involving the carboxyl group and the phenolic hydroxyl group at the C7 center of 3 f (see 3 g in Scheme 2) prevailed over the expected thioesterification.…”

Daltonkins A and B, Two New Carboxyethylflavanones from the Heartwood of Dalbergia tonkinensis

“…Up until the present, not many natural compounds with side chains containing propanoic acid moiety or carboxyethyl groups have been discovered. The few reported products include derivatives of phenylpropanoic acid from the heartwood of Cordia trichotoma and a Streptomycetes strain; carboxyethylchromanones from the bark of Calophyllum papuanum and the fruit of Foeniculum vulgare ; and carboxyethylflavonoids as a biosynthetic product from Vitis vinifera leucoanthocyanidin dioxygenase . With regards to species of Dalbergia , only L‐3,4‐dihydroxyphenylalanine (L‐DOPA) had been isolated from the seeds of D. retusa and D. glabra .…”

Immobilization of flavan-3-ols onto sensor chips to study their interactions with proteins and pectins by SPR

“…In order to immobilize flavan-3-ol monomers/oligomers onto sensor chips, a linker bearing a protected thiol group at one of its extremities was synthesized. The other extremity of the linker was bound to the flavan-3-ol catecholic B-ring through a Vilarrasa reaction [23,26]. In order to optimize the mobility of flavan-3-ols immobilized onto surfaces, flavan-3-ol catechols were modified by using only one equivalent of the linker per equivalent of flavan-3-ol.…”