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| Author(s): |
Amadeu Hoshi Iglesias
Total Authors: 1
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| Document type: | Doctoral Thesis |
| Press: | Campinas, SP. |
| Institution: | Universidade Estadual de Campinas (UNICAMP). Instituto de Química |
| Defense date: | 2009-03-04 |
| Examining board members: |
Fábio Cesar Gozzo;
Luiz Alberto Beraldo de Moraes;
Emer Suavinho Ferro;
Carlos Henrique Inacio Ramos;
Munir Salomão Skaf
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| Advisor: | Fábio Cesar Gozzo |
| Abstract | |
Mass Spectrometry (MS) is the most important tool for analyses related to protein primary structure. Cross-Linking and footprinting aim to bring those advantages to gain insights in the spatial structure of protein complexes. In this work the fragmentation of peptides containing DSS, the most used cross-linker, was studied. Fragmentation mechanisms were proposed based on the dissociation of model peptides which were synthesized in order to generate species that would resemble species found in experiments with proteins. Diagnostic ions were identified which allowed the use of Precursor Ion Scan to detect those species. In order to perform footprinting experiments, a new beamline was developed at the LNLS. A new method for quantitation of oxidation kinetics was developed based on LC-MS analysis, which considers every single oxidation product that can be generated. Those methods were thereafter used in the study of the interaction between Tif34 and Tif35, proteins which compose a translation initiation factor of Saccharomyces cerevisiae. The results indicate that Tif34 presents two different sites for the interaction with Tif35 (AU) | |