Arabidopsis methyltransferase fingerprints by affinity-based protein profiling.
Precise annotation of time and spatial distribution of enzymes involved in plant secondary metabolism by gel electrophoresis are usually difficult due to their low abundance. Therefore, effective methods to enrich these enzymes are required to correlate available transcript and metabolite data with the actual presence of active enzymes in wild-type and mutant plants or to monitor variations of these enzymes under various types of biotic and abiotic stress conditions. S-Adenosyl-L-methionine-dependent O-methyltransferases play important roles in the modification of natural products such as phenylpropanoids or alkaloids. In plants they occur as small superfamilies with defined roles for each of its members in different organs and tissues. We explored the use of S-adenosyl-L-homocysteine as a selectivity function in affinity-based protein profiling supported by capture compound mass spectrometry. Due to their high affinity to this ligand it was possible to identify developmental changes of flower-specific patterns of plant natural product O-methyltransferases and corroborate the absence of individual O-methyltransferases in the corresponding Arabidopsis knockout lines. Developmental changes in the OMT pattern were correlated with transcript data obtained by qPCR.
Wirsing
Lisette
L
Leibniz Institute of Plant Biochemistry, Department of Secondary Metabolism, Weinberg 3, D-06120 Halle (Saale), Germany.
Naumann
Kai
K
Vogt
Thomas
T
D017360:Arabidopsis / Q000201:enzymology*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D013058:Mass Spectrometry / Q000379:methods*; D008780:Methyltransferases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D009154:Mutation; D010455:Peptides / Q000032:analysis; D012435:S-Adenosylhomocysteine / Q000737:chemistry; D014357:Trypsin / Q000378:metabolism
D010455:Peptides; D012435:S-Adenosylhomocysteine; D008780:Methyltransferases; D014357:Trypsin