A radioassay of enzymes catalyzing the glucosylation and sulfation steps of glucosinolate biosynthesis in Brassica species.
A new method for assaying the enzymes uridine diphosphoglucose (UDPglucose):thiohydroximate glucosyltransferase and 3'-phosphoadenosine-5'-phosphosulfate:desulfoglucosinolate sulfotransferase has been designed. The assay system is based on the separation of nonionic [14C]desulfobenzylglucosinolate from anionic [14C]UDPglucose and anionic [14C]benzylglucosinolate, respectively, by differential adsorption to DEAE-ion-exchange disks. The procedure eliminates elaborate chromatographic techniques. The method was used to measure both enzymes in several Brassica spp. In addition, sulfotransferase activity was monitored during partial purification from seedlings of Brassica napus (cv Westar).
Jain
J C
JC
Plant Biotechnology Institute, National Research Council, Saskatoon, Saskatchewan, Canada.
Reed
D W
DW
GrootWassink
J W
JW
Underhill
E W
EW
D001937:Brassica / Q000201:enzymology; D002850:Chromatography, Gel; D005961:Glucosinolates / Q000096:biosynthesis*; D005964:Glucosyltransferases / Q000032:analysis* / Q000302:isolation & purification; D011869:Radioligand Assay; D015238:Sulfotransferases / Q000032:analysis* / Q000302:isolation & purification; D013865:Thioglycosides / Q000096:biosynthesis*; D013997:Time Factors; D014532:Uridine Diphosphate Glucose / Q000032:analysis*; D014539:Uridine Diphosphate Sugars / Q000032:analysis*
D005961:Glucosinolates; D013865:Thioglycosides; D014539:Uridine Diphosphate Sugars; D005964:Glucosyltransferases; C059880:UPD-glucose-thiohydroximate glucosyltransferase; D015238:Sulfotransferases; C059883:desulfoglucosinolate sulfotransferase; D014532:Uridine Diphosphate Glucose