Project TE
Project information
Source of funding : National Council of Scientific Research in Higher Education – CNCSIS, UEFSCDI.
Project Code: PN-II-RU-TE-2011-3-0038.
Number of contract : 112 / 31.10.2011
Project title : New Mass-Spectrometric-based Approaches to Elucidation of Oxidative Modifications in Proteins
Project description
Scientific context and motivation. The nitration of tyrosine residues in protein represents an important post-translational modification during development, oxidative stress, and biological aging; however it is difficult to be detected specifically. Nitration is a covalent modification in proteins resulting from the addition of a nitro group onto one of the two equivalent aromatic carbons adjacent to the hydroxyl group of a tyrosine residue. This modification leads to changes in protein structure and function. Only a selective number of proteins are modified by nitration in vivo, and the selectivity may be influenced by a combination of several factors such as (1), the proteins are in close proximity to the site of generation of nitrating agents; (2), the chemical selectivity of the nitrating reagent; (3), the relative abundance of the target proteins; (4), the proteins contain tyrosine residues in a specific primary sequence or in a specific environment and (5), the repair of nitrated proteins by a putative enzyme called denitrase [1].
Objectives
The scientific objectives of the proposed project comprised two major parts:
I. Affinity-mass spectrometric based approaches to the identification of tyrosine nitration in sputum samples of patients and purified proteins.
Since the identification of nitration sites in cellular proteins have been identified only in a few cases, and structural details have not been characterised in the literature, the application of the new developed affinity-mass spectrometry will provide important results which can be used further in claryfing and understanding the molecular basis of pathophysiological biochemistry. New nitrated target proteins and new nitration sites will be identified and their specific and selective properties will be evaluated.
Scientific reports
Noi abordari mass-spectrometrice pentru elucidarea modificarilor oxidative în protein”
În concordanţă cu obiectivele proiectului stabilite iniţial o mare majoritate a acestora au fost realizate pana in prezent.
In prima etapa a proiectului (Etapa I) s-a realizat perioada de documentare și pregatire a literaturii pentru a fi pusă la dispoziţia studenţilor ce au fost cooptaţi în echipă. Am pregătit materiale de studiu ale unor technici relativ nou introduse în laboratoarele biochimice si cele de chimie analitică, pe care le folosim în cadrul proiectului.
Aceste technici analitice constituie baza unei strategii de cercetare numită proteomica. Proteomica reprezintă un domeniu nou de cercetare folosit cu succes în identificarea proteinelor din probe complexe, determinând structura, modificarile lor post-translationale cat si relatiile dintre proteine si liganzi. Proteomica foloseşte tehnologii ce pot analiza cu succes sute de proteine într-un singur experiment.
Dissemination of results
Dissemination of results
Publications in which the financial support of the project was acknowledged:
Journal articles:
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Petre B.A., Ulrich M., Stumbaum M., Bernevic ., Moise A., Döring G., Przybylski M. (2012), When is Mass Spectrometry Combined with Affinity Approaches Essential? A Case Study of Tyrosine Nitration in Proteins, J. Am. Soc. Mass Spectrom, 23(11): 1831-1840.
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Schreier V. N., Pethő L, Orbán E., Marquardt A., Petre B.A., Mező G. and Manea M. (2014), Protein expression profile of HT-29 human colon cancer cells after treatment with a cytotoxic daunorubicin-GnRH-III derivative bioconjugate, PLOS ONE – in press