Neprosin, a Selective Prolyl Endoprotease for Bottom-up Proteomics and Histone Mapping
The result's identifiers
Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61388971%3A_____%2F17%3A00477176" target="_blank" >RIV/61388971:_____/17:00477176 - isvavai.cz</a>
Alternative codes found
RIV/00216208:11310/17:10364670
Result on the web
<a href="http://dx.doi.org/10.1074/mcp.M116.066803" target="_blank" >http://dx.doi.org/10.1074/mcp.M116.066803</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1074/mcp.M116.066803" target="_blank" >10.1074/mcp.M116.066803</a>
Alternative languages
Result language
angličtina
Original language name
Neprosin, a Selective Prolyl Endoprotease for Bottom-up Proteomics and Histone Mapping
Original language description
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 end point, resulting in an average peptide mass of 2521 units and a higher dependence upon electron-transfer dissociation for peptide-spectrum matches. In contrast to most proline-cleaving enzymes, neprosin effectively degrades proteins of any size. For 1251 HeLa cell proteins identified in common using trypsin, Lys-C, and neprosin, almost 50% of the neprosin sequence contribution is unique. The high average peptide mass coupled with cleavage at residues not usually modified provide new opportunities for profiling clusters of post-translational modifications. We show that neprosin is a useful reagent for reading epigenetic marks on histones. It generates peptide 1-38 of histone H3 and peptide 1-32 of histone H4 in a single digest, permitting the analysis of co-occurring post-translational modifications in these important N-terminal tails.
Czech name
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Czech description
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Classification
Type
J<sub>imp</sub> - Article in a specialist periodical, which is included in the Web of Science database
CEP classification
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OECD FORD branch
10608 - Biochemistry and molecular biology
Result continuities
Project
Result was created during the realization of more than one project. More information in the Projects tab.
Continuities
I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace
Others
Publication year
2017
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data specific for result type
Name of the periodical
Molecular and Cellular Proteomics
ISSN
1535-9476
e-ISSN
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Volume of the periodical
16
Issue of the periodical within the volume
6
Country of publishing house
US - UNITED STATES
Number of pages
10
Pages from-to
1162-1171
UT code for WoS article
000402576600015
EID of the result in the Scopus database
2-s2.0-85020434004