Next Article in Journal
Short-Term Neonatal Oral Administration of Oleanolic Acid Protects against Fructose-Induced Oxidative Stress in the Skeletal Muscles of Suckling Rats
Next Article in Special Issue
Anti-Idiotype DNA Aptamer Affinity Purification–High-Temperature Reversed-Phase Liquid Chromatography: A Simple, Accurate, and Selective Bioanalysis of Bevacizumab
Previous Article in Journal
Graphene-Based Nanocomposites for Neural Tissue Engineering
Previous Article in Special Issue
Simultaneous Detection of Carnosine and Anserine by UHPLC-MS/MS and Its Application on Biomarker Analysis for Differentiation of Meat and Bone Meal
Article Menu
Issue 4 (February-2) cover image

Export Article

Open AccessArticle
Molecules 2019, 24(4), 659; https://doi.org/10.3390/molecules24040659

Optimisation of Protein Extraction from Medicinal Cannabis Mature Buds for Bottom-Up Proteomics

Agriculture Victoria Research, AgriBio, Centre for AgriBioscience, Bundoora, Victoria 3083, Australia
*
Author to whom correspondence should be addressed.
Academic Editor: Makoto Tsunoda
Received: 25 January 2019 / Revised: 11 February 2019 / Accepted: 11 February 2019 / Published: 13 February 2019
(This article belongs to the Special Issue Biological Sample Analysis by Liquid Chromatography)
Full-Text   |   PDF [3835 KB, uploaded 13 February 2019]   |  

Abstract

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 we know, no optimisation of protein extraction from cannabis reproductive tissues has been 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 protein sequences from C. sativa and closely related species 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 nano-liquid chromatography tandem mass spectrometry (nLC-MS/MS) analyses. This is validated by focusing on enzymes involved in the phytocannabinoid pathway. View Full-Text
Keywords: urea; guanidine-HCl; trypsin digestion; nLC-MS/MS; apical bud; trichome urea; guanidine-HCl; trypsin digestion; nLC-MS/MS; apical bud; trichome
Figures

Figure 1

This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).

Supplementary material

SciFeed

Share & Cite This Article

MDPI and ACS Style

Vincent, D.; Rochfort, S.; Spangenberg, G. Optimisation of Protein Extraction from Medicinal Cannabis Mature Buds for Bottom-Up Proteomics. Molecules 2019, 24, 659.

Show more citation formats Show less citations formats

Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Related Articles

Article Metrics

Article Access Statistics

1

Comments

[Return to top]
Molecules EISSN 1420-3049 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert
Back to Top