Identification of a novel flavonoid glycoside sulfotransferase in Arabidopsis thaliana.
The discovery of sulfated flavonoids in plants suggests that sulfation may play a regulatory role in the physiological functions of flavonoids. Sulfation of flavonoids is mediated by cytosolic sulfotransferases (SULTs), which utilize 3'-phosphoadenosine 5'-phosphosulfate (PAPS) as the sulfate donor. A novel SULT from Arabidopsis thaliana, designated AtSULT202B7 (AGI code: At1g13420), was cloned and expressed in Escherichia coli. Using various compounds as potential substrates, we demonstrated, for the first time, that AtSULT202B7 displayed sulfating activity specific for flavonoids. Intriguingly, the recombinant enzyme preferred flavonoid glycosides (e.g. kaempferol-3-glucoside and quercetin-3-glucoside) rather than their aglycone counterparts. Among a series of hydroxyflavones tested, AtSULT202B7 showed the enzymatic activity only for 7-hydroxyflavone. pH-dependency study showed that the optimum pH was relatively low (pH 5.5) compared with those (pH 6.0-8.5) previously reported for other isoforms. Based on the comparison of high performance (pressure) liquid chromatography (HPLC) retention times between sulfated kaempferol and the deglycosylated product of sulfated kaempferol-3-glucoside, the sulfation site in sulfated kaempferol-3-glucoside appeared to be the hydroxyl group of the flavonoid skeleton. In addition, by using direct infusion mass spectrometry, it was found that the sulfated product had one sulfonate group within the molecule. These results indicated that AtSULT202B7 functions as a flavonoid glycoside 7-sulfotransferase.
['Hashiguchi|Takuyu|T|', 'Sakakibara|Yoichi|Y|', 'Shimohira|Takehiko|T|', 'Kurogi|Katsuhisa|K|', 'Yamasaki|Masao|M|', 'Nishiyama|Kazuo|K|', 'Akashi|Ryo|R|', 'Liu|Ming-Cheh|MC|', 'Suiko|Masahito|M|']
D000595:Amino Acid Sequence D017360:Arabidopsis D015239:Arylsulfotransferase D002851:Chromatography, High Pressure Liquid D004591:Electrophoresis, Polyacrylamide Gel D006863:Hydrogen-Ion Concentration D044949:Kaempferols D008969:Molecular Sequence Data D009005:Monosaccharides D013379:Substrate Specificity
flavonoid glycoside sulfotransferase Arabidopsis thaliana sulfation AtSULT202B7