Substrate profiling of cysteine proteases using a combinatorial peptide library identifies functionally unique specificities

The Journal of Biological Chemistry
Youngchool ChoeCharles S Craik

Abstract

The substrate specificities of papain-like cysteine proteases (clan CA, family C1) papain, bromelain, and human cathepsins L, V, K, S, F, B, and five proteases of parasitic origin were studied using a completely diversified positional scanning synthetic combinatorial library. A bifunctional coumarin fluorophore was used that facilitated synthesis of the library and individual peptide substrates. The library has a total of 160,000 tetrapeptide substrate sequences completely randomizing each of the P1, P2, P3, and P4 positions with 20 amino acids. A microtiter plate assay format permitted a rapid determination of the specificity profile of each enzyme. Individual peptide substrates were then synthesized and tested for a quantitative determination of the specificity of the human cathepsins. Despite the conserved three-dimensional structure and similar substrate specificity of the enzymes studied, distinct amino acid preferences that differentiate each enzyme were identified. The specificities of cathepsins K and S partially match the cleavage site sequences in their physiological substrates. Capitalizing on its unique preference for proline and glycine at the P2 and P3 positions, respectively, selective substrates and a substrate-...Continue Reading

References

Feb 12, 1970·Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences·A Berger, I Schechter
Jun 10, 1994·Journal of Medicinal Chemistry·B M WagnerA Krantz
May 1, 1994·Biological Chemistry Hoppe-Seyler·D BrömmeA Krantz
Apr 1, 1996·Protein Science : a Publication of the Protein Society·D Brömme, M E McGrath
Jul 9, 1998·Protein Science : a Publication of the Protein Society·M E McGrathJ R Somoza
Dec 9, 1998·Proceedings of the National Academy of Sciences of the United States of America·M B EisenD Botstein
Mar 10, 2000·Biochimica Et Biophysica Acta·B TurkV Turk
Jun 28, 2000·Proceedings of the National Academy of Sciences of the United States of America·J L HarrisC S Craik
Jul 5, 2001·The Journal of Biological Chemistry·J L HarrisC S Craik
Sep 5, 2001·The EMBO Journal·V TurkD Turk
Feb 19, 2002·Molecular and Biochemical Parasitology·M Sajid, J H McKerrow
May 3, 2002·The Journal of Biological Chemistry·Jason P SalterJames H McKerrow
Jul 23, 2002·Current Pharmaceutical Design·Dieter Brömme, Jadwiga Kaleta
Dec 19, 2003·Nucleic Acids Research·Neil D RawlingsAlan J Barrett
Sep 17, 2004·Archives of Biochemistry and Biophysics·Luciano PuzerAdriana K Carmona
Dec 8, 2004·International Journal for Parasitology·Philip J Rosenthal
Jan 22, 2005·Trends in Biotechnology·Alan B Marnett, Charles S Craik
May 7, 2005·Advanced Drug Delivery Reviews·Yoshiyuki YasudaDieter Brömme

❮ Previous
Next ❯

Citations

Mar 4, 2010·Glycoconjugate Journal·Claudia PönighausChristian Götting
Jul 10, 2013·Journal of Chemical Information and Modeling·Serghei Glinca, Gerhard Klebe
Aug 20, 2009·Journal of the American Chemical Society·Xu Wu, Peter G Schultz
Jun 13, 2012·Journal of Medicinal Chemistry·Maxim FrizlerMichael Gütschow
Apr 25, 2013·Journal of Medicinal Chemistry·Roberta EttariPaola Conti
Oct 20, 2007·Nature Chemical Biology·Kerry Routenberg LoveHidde L Ploegh
Feb 18, 2011·Nature Communications·Melanie A Adams-CioabaJinrong Min
Oct 2, 2012·Nature Methods·Anthony J O'DonoghueCharles S Craik
Sep 3, 2010·Nature Reviews. Drug Discovery·Marcin Drag, Guy S Salvesen
Jul 22, 2009·The Journal of Biological Chemistry·Iain D KerrLinda S Brinen
Oct 22, 2010·The Journal of Biological Chemistry·Ryan K SwenertonJames H McKerrow
Aug 13, 2011·The Journal of Biological Chemistry·Adéla JílkováMichael Mareš
May 1, 2008·The Journal of Biological Chemistry·Kelly N DuBoisMohammed Sajid
Aug 15, 2008·The Journal of Biological Chemistry·Theresa C O'BrienJames H McKerrow
Oct 30, 2008·The Journal of Biological Chemistry·Nicolas Lützner, Hubert Kalbacher
Dec 6, 2011·Briefings in Bioinformatics·David A duVerle, Hiroshi Mamitsuka
Nov 15, 2011·Journal of Clinical Microbiology·Wendy E KamanFloris J Bikker
May 3, 2013·PLoS Computational Biology·Julian E FuchsKlaus R Liedl
Jun 3, 2009·PLoS Neglected Tropical Diseases·Jan DvorákConor R Caffrey
Jan 23, 2008·Biological Chemistry·Petra Van DammeKris Gevaert
Aug 12, 2009·Biological Chemistry·Rodrigo L O R CunhaLuiz Juliano
Jun 2, 2012·The Journal of Endocrinology·Marcela OliveiraAdriana K Carmona
Oct 7, 2014·Biochemistry·Russell F Doolittle

❮ Previous
Next ❯

Related Concepts

Related Feeds

ASBMB Publications

The American Society for Biochemistry and Molecular Biology (ASBMB) includes the Journal of Biological Chemistry, Molecular & Cellular Proteomics, and the Journal of Lipid Research. Discover the latest research from ASBMB here.