It did not transfer a methyl group to any hydroxyl other than with the 7 or 4# place. Radioactive TLC of the response with myricetin revealed PD98059 selleckchem a single item that migrated between myricetin and 3# methyl myricetin. This merchandise was identified by LC MS as seven methyl myricetin. When myricetin was incubated with ShMOMT2 overnight, the solution obtained was 7,4# methyl myricetin. The Structural Relatedness of ShMOMT1 and ShMOMT2 to Other OMTs The protein encoded by the ShMOMT1 cDNA is 362 amino acids extended, using a calculated molecular mass of 40.7 kD, and it is made up of all the recognized plant OMT domains known or hypothesized to get involved in binding to SAM and metal cofactors. ShMOMT1 is most related to a lot of mainly 3# and 3#/5# OMTs, steady with its regiospecificity for the 3# and 5# positions. The protein encoded through the ShMOMT2 cDNA is 355 amino acids long, by using a calculated molecular mass of 39.4 kD, and additionally, it contains all the recognized plant OMT domains regarded or hypothesized to become concerned in binding to SAM and metal cofactors. ShMOMT2 ismost very similar to a few OMTs identified as specific for your seven and/or 4# place, steady with its regiospecificity for these positions.
ShMOMT2 is only 27% identical to ShMOMT1. Distribution of ShMOMT1 and ShMOMT2 Transcripts and Protein in Trichome Glands We utilized quantitative reverse transcription PCR and western blot analyses Phlorizin to localize ShMOMT1 and ShMOMT2 transcripts, and ShMOMT1 and ShMOMT2 proteins, respectively, in the various kinds of trichome glands. Extracts of collections of personal kinds of glands have been compared with full leaf extracts in the two kinds of experiments. ShMOMT1 transcript ranges had been three.five to 12.5 fold greater in secreting glands from styles 4 and 1 trichomes, respectively, compared with storage glands of variety six trichomes. ShMOMT2 transcript ranges have been 2 to 4 fold larger in secreting glands from varieties 4 and one trichomes, respectively, compared with storage glands of sort six trichomes. Comparison of transcript amounts from leaf tissue with trichomes versus leaf tissue from which the trichomes had been mechanically removed indicated that transcripts of each ShMOMT1 and ShMOMT2 are current exclusively in trichomes. Protein blot evaluation indicated that levels of ShMOMT1 protein were seven to 8.six fold greater in secreting glands compared with storage glands and one.9 to 2.four fold greater in contrast with total leaf extracts. Levels of ShMOMT2 protein had been 5 to six.six fold higher in secreting glands compared with storage glands, though ShMOMT2 was not detectable in whole leaf extracts.