A Protease for Middle Down Proteomics

We developed a method for restricted enzymatic proteolysis using the outer membrane protease T (OmpT) to produce large peptides (> 6.3 kDa on average) for mass spectrometry-based proteomics. Using this approach to analyze prefractionated high-mass HeLa proteins we identified 3,697 unique peptides...

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Detalles Bibliográficos
Autores principales: Wu, Cong, Tran, John C., Zamdborg, Leonid, Durbin, Kenneth R., Li, Mingxi, Ahlf, Dorothy R., Early, Bryan P., Thomas, Paul M., Sweedler, Jonathan V., Kelleher, Neil L.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: 2012
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3430368/
https://www.ncbi.nlm.nih.gov/pubmed/22706673
http://dx.doi.org/10.1038/nmeth.2074
Descripción
Sumario:We developed a method for restricted enzymatic proteolysis using the outer membrane protease T (OmpT) to produce large peptides (> 6.3 kDa on average) for mass spectrometry-based proteomics. Using this approach to analyze prefractionated high-mass HeLa proteins we identified 3,697 unique peptides from 1,038 proteins. We demonstrated the ability of large OmpT peptides to differentiate closely related protein isoforms and to enable the detection of many post-translational modifications.

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