Curso 3 - Rosa

Curso  3: Advanced Mass Spectrometry-based proteomics for Post-Translational Modifications (PTMs) Analysis
Responsável: Prof. José Cesar Rosa, FMRP-USP e Prof. Giuseppe Palmisano, IB-USP

Cellular signalling is a fundamental biomolecular response of biological systems to biotic or abiotic stimuli. The activation of signalling events is mediated by protein post-translational modifications (PTMs) such as phosphorylation, acetylation, ubiquitination, glycosylation, oxidation and other understudied modifications. These modifications functionally decorate the protein primary sequence changing their structure and function. The dysregulation of specific PTMs has been associated to disease such as cancer, diabetes, cardiovascular, neurodegenerative, infectious and others. Indeed, several drug targets are enzymes that catalyse the addition (e.g. kinases) or removal (e.g. sirtuins) of PTMs. Moreover, modified proteins/peptides can be used as a biomarker treasure to achieve high selectivity and specificity. Recently, a new paradigm has been proposed regarding PTMs crosstalk. Despite the knowledge of highly dynamic and concerted regulation of PTMs in histone tails, it is recognized nowadays that PTMs regulate the function of enzymes involved in other PTMs biosynthesis. Overall, it has been demonstrated that PTMs acts in concert and their expression constitute the PTM code.
Mass spectrometry is the gold standard in proteomics studies. The application of mass spectrometry in proteomics has improved our current understanding of biological systems by identifying and quantifying different PTMs in a system-wide approach. However, the improvement has been possible due to the combination of dedicated sample preparation strategies, such as enrichment methods and advanced computational approaches. Nowadays, researchers in the proteomic field benefit from several tools to study PTMs. These tools are the basis for future developments in the area.
Aims: This course aims at: 1) providing a complementary perspective on the use of mass spectrometry to identify and quantify protein PTMs. Due to the importance of PTMs in several scientific fields, this course will 2) present the state-of-the-art instrumentation and sample preparation for PTMs analysis using mass spectrometry. 3) Offer new possibilities to implement PTMs analysis in the students’ laboratories. 4) Expand the repertoire of tools available in the proteomic field. 5) Highlight the capabilities of mass spectrometry to identify and quantify PTMs in large-scale experiments.
Course organization: The theoretical part of the course will be divided in three main parts: 1) biological significance of PTMs, 2) enrichment methods and 3) mass spectrometry-based proteomics analysis of PTMs. There will be a group discussion that will aim at giving suggestions to improve the field.

After a brief introduction of all the participants, the morning session from 8:30 to 12:00 will be dedicated to lectures on:
1) Biological significance, enrichment methods and mass spectrometry analysis of protein phosphorylation;
2) Biological significance, enrichment methods and mass spectrometry analysis of protein glycosylation;
3) Overview of other understudied PTMs.

Afternoon session from 13:00-16:00h
The afternoon session will be dedicated to discussion of previously selected and relevant publication on PTMs send to participants. This presentation will be an opportunity to present an up-to-date research on the field. During this occasion, will be presented newly developed techniques described in the literature and also each participant will share his/her doubts and challenges with other colleagues. The discussion will bring everyone together in order to develop a task force to solve the problems in PTM analysis.

1) Carga horária: 6h (T).
2) Datas: 27 de julho de 2017, 8:30 às 16:00.
3) Local: Hotel Monte Real, Águas de Lindóia
4) Nº máximo de participantes:
5) Valor do Investimento: R$ 50,00



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