Tryp-N: A Thermostable Protease for the Production of N-terminal Argininyl and Lysinyl Peptides.

Journal of Proteome Research
John P WilsonDarryl J Pappin

Abstract

Bottom-up proteomics is a mainstay in protein identification and analysis. These studies typically employ proteolytic treatment of biological samples to generate suitably sized peptides for tandem mass spectrometric (MS) analysis. In MS, fragmentation of peptides is largely driven by charge localization. Consequently, peptides with basic centers exclusively on their N-termini produce mainly b-ions. Thus, it was long ago realized that proteases that yield such peptides would be valuable proteomic tools for achieving simplified peptide fragmentation patterns and peptide assignment. Work by several groups has identified such proteases, however, structural analysis of these suggested that enzymatic optimization was possible. We therefore endeavored to find enzymes that could provide enhanced activity and versatility while maintaining specificity. Using these previously described proteases as informatic search templates, we discovered and then characterized a thermophilic metalloprotease with N-terminal specificity for arginine and lysine. This enzyme, dubbed Tryp-N, affords many advantages including improved thermostability, solvent and detergent tolerance, and rapid digestion time.

References

Jan 1, 1977·Methods in Enzymology·W M Mitchell
Nov 15, 1984·Analytical Biochemistry·S S Twining
Jan 1, 1995·Methods in Enzymology·N D Rawlings, A J Barrett
Oct 30, 1998·Journal of the American Society for Mass Spectrometry·J P DeGnore, J Qin
Feb 4, 2003·Nature Reviews. Drug Discovery·Gary L Glish, Richard W Vachet
Sep 9, 1961·Nature·D A ELLIS
Mar 23, 2004·Molecular & Cellular Proteomics : MCP·Jesper V OlsenMatthias Mann
Sep 2, 2004·Nature Reviews. Molecular Cell Biology·Hanno Steen, Matthias Mann
Apr 22, 2006·The Journal of Biological Chemistry·Cynthia TallantF Xavier Gomis-Rüth
Nov 14, 2006·Archives of Biochemistry and Biophysics·Raquel García-CastellanosF Xavier Gomis-Rüth
Feb 13, 2007·Nature Methods·Mamie Z Li, Stephen J Elledge
Dec 11, 2007·Journal of Proteome Research·Jesse RodriguezPavel A Pevzner
Feb 7, 2009·Journal of Proteome Research·Laura HohmannDaniel B Martin
Jun 2, 2010·Nucleic Acids Research·Mark N WassMichael J E Sternberg
Jun 15, 2010·Journal of Proteome Research·Nadia TaouatasShabaz Mohammed
Mar 23, 2011·Nature Genetics·Susanna ObadSakari Kauppinen
Dec 3, 2013·Nucleic Acids Research·Antonina AndreevaAlexey G Murzin
Oct 29, 2014·Nucleic Acids Research·UNKNOWN UniProt Consortium
Nov 25, 2014·Nature Methods·Pitter F HuesgenChristopher M Overall
May 8, 2015·Nature Protocols·Lawrence A KelleyMichael J E Sternberg
Jun 5, 2015·ACS Chemical Biology·Giada MarinoChristopher M Overall
Dec 15, 2015·Proteomics·Stéphane TrevisiolBruno Domon
Apr 29, 2016·Nature Protocols·Piero GiansantiAlbert J R Heck
Mar 21, 2017·Methods in Molecular Biology·Fatih DemirPitter F Huesgen
Oct 17, 2017·Bioinformatics·Omar Wagih
May 23, 2018·Nucleic Acids Research·Andrew WaterhouseTorsten Schwede
Oct 24, 2018·Molecular Biology and Evolution·Guangchuang YuYi Guan
Nov 6, 2018·Nucleic Acids Research·Yasset Perez-RiverolJuan Antonio Vizcaíno
Apr 13, 2019·Nucleic Acids Research·Fábio MadeiraRodrigo Lopez
May 9, 2019·Rapid Communications in Mass Spectrometry : RCM·Shu LiuPing Xu
Jun 22, 2019·Science·Dannielle D EngleDavid A Tuveson
Dec 25, 2019·Nature Chemical Biology·Avinash D LondheBenoit Boivin

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