A sensitive and rapid bioassay of homogalacturonan synthase using 2-aminobenzamide-labeled oligogalacturonides.
Polygalacturonate 4-alpha-galacturonosyltransferase (GalA T) activity was detected in the microsomal fraction isolated from pumpkin (Cucurbia moschata Duchesne, cv. Tokyou-Kabocha) seedlings using UDP-GalA and 2-aminobenzamide (2AB)-labeled oligogalacturonides. A 2AB-labeled undecagalacturonide was elongated by the attachment of galacturonic acid (GalA) residues to give 2AB-labeled oligogalacturonides with a degree of polymerization (DP) between 12 and 17. Exogenous 2AB-labeled oligogalacturonide acceptors with a DP >3 are effective acceptor molecules for pumpkin GalA T.
Ishii
Tadashi
T
Forestry and Forest Products Research Institute, P. O. Box 16, Tsukuba Norin Kenkyu Danchi-nai, Ibaraki, 305-8687 Japan. tishii@ffpri.affrc.go.jp
D001681:Biological Assay / Q000379:methods*; D028464:Cucurbita / Q000378:metabolism; D014453:Glucuronosyltransferase; D016695:Glycosyltransferases / Q000378:metabolism*; D008861:Microsomes / Q000378:metabolism; D009844:Oligosaccharides / Q000378:metabolism*; D010940:Plant Proteins / Q000378:metabolism*; D014539:Uridine Diphosphate Sugars / Q000378:metabolism; D062367:ortho-Aminobenzoates / Q000378:metabolism*
D009844:Oligosaccharides; D010940:Plant Proteins; D014539:Uridine Diphosphate Sugars; C412763:oligogalacturonic acid; D062367:ortho-Aminobenzoates; C050306:UDP-galacturonic acid; D016695:Glycosyltransferases; D014453:Glucuronosyltransferase; C438373:alpha-1,4-galacturonosyltransferase; C000219:anthranilamide