Neprosin, a Selective Prolyl Endoprotease for Bottom-up Proteomics and Histone Mapping.

ABSTRACT: Trypsin dominates bottom-up proteomics, but there are reasons to consider alternative enzymes. Improving sequence coverage, exposing proteomic "dark matter," and clustering post-translational modifications in different ways and with higher-order drive the pursuit of reagents complementary to trypsin. Additionally, enzymes that are easy to use and generate larger peptides that capitalize upon newer fragmentation technologies should have a place in proteomics. We expressed and characterized recombinant neprosin, a novel prolyl endoprotease of the DUF239 family, which preferentially cleaves C-terminal to proline residues under highly acidic conditions. Cleavage also occurs C-terminal to alanine with some frequency, but with an intriguingly high "skipping rate." Digestion proceeds to a stable

SUBMITTER: Schrader CU 

PROVIDER: S-EPMC5461545 | biostudies-literature | 2017 Jun

REPOSITORIES: biostudies-literature