RESUMO
The study of protein misfolding and post-translational processing abnormalities is a promising diagnostic approach for socially significant pathologies associated with the accumulation of abnormal forms of proteins. Recently, it was shown that amyloid-like aggregates can be observed in the urine of pregnant women with preeclampsia, which is the most severe hypertensive complication that can lead to fateful outcomes. The protein composition of urine aggregates may clarify the molecular mechanisms underlying the pathology and has not yet been studied in detail. Using a proteomic approach based on high-resolution mass spectrometry, we studied the protein composition of amyloid-like structures that aggregate in the presence of Congo red azo-dye in the urine of pregnant women with preeclampsia. Fragments of ß-sheets of α-1-antitrypsin, complement 3, haptoglobin, ceruloplasmin, and trypstatin were identified as most likely targets for Congo red binding.
Assuntos
Amiloide/urina , Espectrometria de Massas/métodos , Pré-Eclâmpsia/urina , Proteoma/análise , Amiloide/química , Vermelho Congo , Feminino , Humanos , Pré-Eclâmpsia/metabolismo , Gravidez , Proteoma/química , Proteômica/métodosRESUMO
Urine is a sample of choice for noninvasive biomarkers search because it is easily available in large amounts and its molecular composition provides information on processes in the organism. The high potential of urine peptidomics has been demonstrated for clinical purpose. Several mass spectrometry based approaches have been successfully applied for urine peptidome analysis and potential biomarkers search. Summarizing literature data and our own experience we developed a protocol for comprehensive urine peptidome analysis. The technology includes several stages and consists of urine sample preparation by size exclusion chromatography and identification of featured peptides by nano-HPLC coupled to Fourier transform ion cyclotron resonance mass spectrometry, semiquantitative and statistical data analysis.