EndoProteoFASP as a Tool to Unveil the Peptidome-Protease Profile: Application to Salivary Diagnostics.
作者： Fábio Trindade , Inês Falcão-Pires , Adelino Leite-Moreira , Pedro S. Gomes , Julie Klein
DOI: 10.1007/978-1-4939-7537-2_19
关键词:
摘要: In the quest to fully comprehend proteolytic events leading generation of salivary peptidome, we have developed a method for sequential elution peptides throughout progressive endogenous proteolysis. By screening time-dependent changes in peptidome can predict activity pattern proteases responsible such peptide fingerprint and identify susceptible protein targets. Herein, describe step-by-step tutorial based on filter-aided sample preparation (FASP) method, taking advantage armamentarium (endoProteoFASP), produce new from proteins, adding those present at time collection. this protocol, different sets retrieved after are identified following liquid chromatography-tandem mass spectrometry approach. The likelihood large set (collected several public sources) be predicted by analysis cleavage site specificity Proteasix ( http://proteasix.cs.man.ac.uk /) algorithm. attained peptidome-protease profile useful elucidate dynamics underpinning pathophysiological phenomena place locally within oral cavity. This may help clinicians diagnose pathologies develop preventive therapeutic plans.
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