Abstract
Recently we have seen a relaxation of the historic restrictions on the use and subsequent research on the Cannabis plants, generally classified as Cannabis sativa and Cannabis indica. What research has been performed to date has centered on chemical analysis of plant flower products, namely cannabinoids and various terpenes that directly contribute to phenotypic characteristics of the female flowers. In addition, we have seen many groups recently completing genetic profiles of various plants of commercial value. To date, no comprehensive attempt has been made to profile the proteomes of these plants. We report herein our progress on constructing a comprehensive draft map of the Cannabis proteome. To date we have identified over 17,000 potential protein sequences. Unfortunately, no annotated genome of Cannabis plants currently exists. We present a method by which “next generation” DNA sequencing output and shotgun proteomics data can be combined to produce annotated FASTA files, bypassing the need for annotated genetic information altogether in traditional proteomics workflows. The resulting material represents the first comprehensive annotated protein FASTA for any Cannabis plant. Using this annotated database as reference we can refine our protein identifications, resulting in the confident identification of 13,000 proteins with putative function. Furthermore, we demonstrate that post-translational modifications play an important role in the proteomes of Cannabis flower, particularly lysine acetylation and protein glycosylation. To facilitate the evolution of analytical investigations into these plant materials, we have created a portal to host resources developed from our proteomic and metabolomic analysis of Cannabis plant material as well as our results integrating these resources.
Highlights
Proteomics is a science dedicated to the creation of comprehensive quantitative snapshots of all the proteins produced by an individual organism, tissue or cell [1]
A recent quantitative study of Arabidopsis plant material using similar instrumentation as employed here demonstrated an identification rate of approximately 22% against the manually curated UniProt database for this plant. This result is unsurprising given the body of work that has been assembled for this model organism and suggests that further work will be necessary to refine our genomic and proteomic tools for the less studied Cannabis plant [13]
Traditional proteomics workflows rely on the existence of annotated theoretical protein FASTA files derived from annotated genomes
Summary
Proteomics is a science dedicated to the creation of comprehensive quantitative snapshots of all the proteins produced by an individual organism, tissue or cell [1]. While the technology was in place to complete the human genome draft in 2003, the first two drafts of the human proteome were not completed by teams led by Johns Hopkins and Center for Integrated Protein Science Munich researchers until 2014. These two separate and ambitious projects were the composite of thousands of hours of instrument run time utilizing the most sophisticated hardware available at that time [3,4]. Protein modifications are directly involved in nearly every known disease and these modifications are impossible to identify with any current DNA/RNA sequencing technology [11]
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