The information obtained with the powerful Mass Spectrometry techniques is fundamental for the structural characterization of chemical and biochemical species.
Ana Maria Varela Coelho
Phone (+351) 214469451/52/07
The mass spectrometry methods (MS) developed and implemented in our Laboratory allowed several successes in the research work in progress.
The knowledge achieved during the characterization of sea urchin’s adhesive secretions and starfish nerve proteome could be further used in biomimeting biocompatible water-resistent glues and contributing to understand echinoderms impressive regeneration abilities, with likely biomedical applications. Optimization of an efficient solubilization buffer overcame sea-urchin adhesive insolubility enabling the visualization of 13 protein bands by SDS-PAGE. Seven were identified by conjugation of MS with homology-database search, while the remaining 6 seem to be either novel or highly-modified proteins. Several starfish radial nerve cord proteins related with nervous system function and visual perception were identified in echinoderms for the first time, allowing the elucidation of function and mechanisms of this organ in starfish and showing similarities with higher animals synapse mechanisms.
Studies on protein complexes characterization have included optimization of a disassembly procedure and of a MS method for determination of viral proteins molecular mass in viral particles. Results of comparative protein expression profiles investigated in rabbit muscle under food restriction shown relevance of actin in preserving muscle structure and sensitivity of both myosin light chain and α-crystallin protein. Differential proteomic studies demonstrate that an α-amylase isoform is overexpressed in mice saliva from animals fed on tannin rich diets. Observed marked differences between parotid salivary protein profiles of goat and sheep, can be used as a starting point to investigate a relation-ship between physiological function and different feeding behavior of two domesticated species. A study on changes in hepatome proteome induced by hepatitis delta virus replication allowed the identification of proteins involved in virus replication and cellular defence against viral infection. MALDI-TOF data used for the structural characterization of enolase glycation (PTM related with amyloid diseases and diabetes) were correlated with thermal stability and enzyme activity results. These are examples of established and fruitful collaborations with Institutions participating in the National Mass Spectrometry Network, UNL Institutions, other Academic Institutions and Laboratórios do Estado without forgetting the LAO nd international collaborations.
- Peter Boross, Post-doc
- Ricardo Anjos Gomes, Post-doc
- Renata Soares, Post-doc
- Romana Santos, Post-doc
- André Almeida, Post-doc
- Duarte Toubarro, PhD student
- Patrícia Alves, PhD student
- Sérgio Mota, PhD student
- Elsa Lamy, PhD student
- Catarina Franco, PhD student
- Mariana Carvalho, Master student
- Elisabete Pires, Technician
- Conceição Almeida, Technician
- Roxo-Rosa M., da Costa G., Luider T. M., Scholte B. J., Coelho A. V., Amaral M. D. and Penque D. (2006). “Proteomic analysis of nasal cells from cystic fi brosis patients and non-cystic fi brosis control individuals: Search for novel biomarkers of cystic fi brosis lung disease.” Proteomics 6(7): 2314-2325
- Gomes R. A., Miranda H. V., Silva M. S., Graca G., Coelho A. V., Ferreira A. E., Cordeiro C. and Freire A. P. (2006). “Yeast protein glycation in vivo by methylglyoxal - Molecular modifi cation of glycolytic enzymes and heat shock proteins.” FEBS Journal 273(23): 5273-5287.
- Bravo M. N., Silva S., Coelho A. V., Boas L. V. and Bronze M. R. (2006). “Analysis of phenolic compounds in Muscatel wines produced in Portugal.”Analytica Chimica Acta 563(1-2): 84-92
For further information please visit the laboratory's website
A espectrometria de massa (MS) é uma metodologia analítica que permite determinar com exactidão a massa molecular de uma grande variedade de compostos químicos e bioquímicos e obter informação relevante para a sua caracterização estrutural. Nos projectos em curso no Lab. MS usamos esta metodologia para apoiar o desenvolvimento de novos fármacos em colaboração com a indústria farmacêutica e identificar e caracterizar proteínas presentes em células/ tecidos em condições de stress, infecção ou terapia, com o objectivo de aumentar o conhecimento nos processos biológicos em estudo e identificar biomarcadores a usar em terapia e diagnóstico de situações patológicas. Constituem exemplos, os estudos com células infectadas com o vírus da hepatite D e de exsudado nasal de doentes com fibrose quística, a caracterização proteica do músculo e da saliva de animais domésticos e a sua relação com o tipo de dieta administrada. Foram também identificadas novas proteínas adesivas e envolvidas na regeneração do nervo, isoladas de animais marinhos e com aplicações biotecnológicas.