Medicinal cannabis is used to relieve the symptoms of certain medical conditions, such as epilepsy. Cannabis is a controlled substance and until recently was illegal in many jurisdictions. Consequently, the study of this plant has been restricted. Proteomics studies on Cannabis sativa reported so far have been primarily based on plant organs and tissues other than buds, such as roots, hypocotyl, leaves, hempseeds, and flour. As far as our knowledge, no optimisation of protein extraction from cannabis reproductive tissues was attempted. Therefore, we set out to assess different protein extraction methods followed by mass spectrometry-based proteomics to recover, separate and identify the proteins of the reproductive organs of medicinal cannabis, apical buds and isolated trichomes. Database search following shotgun proteomics was limited to C. sativa protein sequences available from UniprotKB. Our results demonstrate that a buffer containing the chaotrope reagent guanidine-hydrochloride recovers many more proteins than a urea-based buffer. In combination with a precipitation with trichloroacetic acid, such buffer proved optimum to identify proteins using a trypsin digestion followed by nLC-MS/MS analyses. This is validated by focusing on enzymes involved in the cannabinoid pathway. doi:10.3390/molecules24040659
[doi:10.25345/C5HG7N]
[dataset license: CC0 1.0 Universal (CC0 1.0)]
Keywords: urea, guanidine-HCl, trypsin digestion, nLC-MS/MS, apical bud, trichome
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Delphine Vincent, Agriculture Victoria Research, Australia |
Submitting User: | delphine_1_2 |
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