Understanding temperature-related physiological processes through proteomics in vegetal and animal systems
Author(s)
Egidi, Maria Giulia
Date Issued
February 8, 2011
Type
Doctoral Thesis
Abstract
Proteomics, the large-scale study of proteins present in a cell, was born and developed with the
basic aim of broaden the current knowledge about the relationship between proteins and the main
biological processes presiding over life of organisms. Due to its instrinsic ability to observe all
protein variations in a biological system in response to different stimuli at once, proteomics could
be resembled to a translator, which converts the language of nature in a network of protein
interactions. The main effort consists in the assembly of the information here obtained to discover
the main factors responsible for these protein variations. The present work describes several
proteomic applications to the characterization of temperature-related phenomenons. Biological
systems, belonging both to vegetal and animal kingdom, were subjected to diverse treatments and
their protein modulations were observed in function of time.
The first objective of my PhD was the study of the influence of prolonged cold treatment on the
expression of cold-induced proteins in a vernalized common wheat (Triticum aestivum L.), to
identify proteomic changes in vernalization-treated plants when the key steps in controlling the
flowering time occur. Leaf proteome of cold acclimated and vernalized winter wheat were
compared with the control non acclimated ones grown at control temperature of 20 °C. Total
protein was extracted from the leaves of cold treated or control plants, then separated by 2-
dimensional electrophoresis. The identities of differentially expressed proteins were determined by
tandem mass spectrometry.
My second objective was the application of proteomics to delve into plant cross-tolerance to
different abiotic stresses. In particular, we focussed on cold and drought stressors; with the attempt
to shed light into those metabolic modulations strictly related to exposure to low temperatures, we
choose a drought-resistant spring wheat variety for our proteomic study so to minimize the
dehydrative component of cold stress.
Finally, in the last year of my PhD, I joined a project concerning platelet quality assessment
through proteomic approach. The aim of my work was to analyze proteomic changes occurring in
platelet concentrates (PCs), in function of time. We used proteomic approach to detect changes in
PC supernatants (obtained after brief centrifugation of PCs) after 0, 4 and 7 days of storage at
22°C. To achieve a more complete overview of the pr otein pool accumulating/dissapearing in this
sub-proteome, we made use of ProteoMinerTM technology to enrich our final proteome for the lowabundance
species, often hidden by high abundant ones (in our case mainly plasma albumin).
The commonality of such different topics was double. First, proteomic approach was applied
almost in the same manner (with proper but minor modifications in the extraction protocols);
second, the general attempt was to achieve a greater understanding of different processes
influenced by temperature through their monitoring over time.
Additional information
Dottorato di ricerca in Genetica e biologia cellulare
Subjects
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