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.

II. Mass-spectrometric based approaches for developing structural characterization of protein aggregates.

The proposed research will develop methods and generate data used to study the structural biology of aggregates using mass spectrometry. Abeta- polypeptides, synuclein and eosinophil cationic protein will be used as model systems to develop new, integrating high performance mass spectrometry – based approaches for (i) chemical structure determination of aggregates; (ii) characterization of interactions in protein aggregation; and (iii) identification of reaction intermediates in aggregate formation, particularly proteolytic fragments and oligomers.