Protein C-terminal modification through thioacid/azide amidation

Bioconjugate Chemistry
Xiaohong ZhangChuan-Fa Liu

Abstract

The preparation of protein bioconjugates has been largely dependent on the development of selective chemistries that are orthogonal to the diverse functionalities present in a protein. Here, we report a new method for C-terminus-directed modification of recombinant proteins. The method is based on the thioacid/azide amidation reaction. Essentially, hydrothiolytic cleavage of the thioester intermediate in protein splicing yields a recombinant protein with a unique thioacid group at the C-terminus, which is then chemoselectively amidated with an electron-poor organic azide carrying a biofunctional tag. The small ubiquitin protein was used as a model system to demonstrate the utility of this new bioconjugation method. C-terminal PEGylation or biotinylation of ubiquitin was readily achieved through amidation of ubiquitin thioacid with a sulfonazide-functionalized PEG or biotin derivative. Our data validate that thioacid/azide amidation is a mechanistically novel and practically useful method for site-selective protein modification.

References

Nov 4, 1994·Science·P E DawsonS B Kent
Jun 17, 1998·Proceedings of the National Academy of Sciences of the United States of America·T W MuirP A Cole
Mar 4, 2003·Nature Reviews. Drug Discovery·J Milton Harris, Robert B Chess
Jun 26, 2003·Journal of the American Chemical Society·Ning ShangguanLawrence J Williams
Aug 19, 2004·Journal of the American Chemical Society·John M Antos, Matthew B Francis
Nov 19, 2004·Chembiochem : a European Journal of Chemical Biology·Alexander DeitersPeter G Schultz
Dec 16, 2004·Angewandte Chemie·Lei Wang, Peter G Schultz
Jan 31, 2006·Chembiochem : a European Journal of Chemical Biology·Dmitry E AgafonovMathias Sprinzl
Feb 24, 2006·Organic Letters·Kristin N BarlettLawrence J Williams
Apr 28, 2006·Journal of the American Chemical Society·Robert V KolakowskiLawrence J Williams
May 16, 2006·Current Opinion in Chemical Biology·John M Antos, Matthew B Francis
Aug 10, 2006·Chembiochem : a European Journal of Chemical Biology·Jeet Kalia, Ronald T Raines
Aug 24, 2006·Nature Reviews. Molecular Cell Biology·Jianming Xie, Peter G Schultz
Dec 27, 2006·Current Opinion in Chemical Biology·Timothy L Foley, Michael D Burkart
Apr 10, 2008·Chembiochem : a European Journal of Chemical Biology·Xiao-Hong TanChuan-Fa Liu

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Citations

Aug 4, 2010·Journal of the American Chemical Society·Kalyanaraman Krishnamoorthy, Tadhg P Begley
Aug 19, 2011·Journal of the American Chemical Society·Wenting WuLanny S Liebeskind
Dec 11, 2013·PloS One·Isabelle SermadirasPeter Ravn
Jun 24, 2014·Journal of Controlled Release : Official Journal of the Controlled Release Society·Xiang YiAlexander V Kabanov
Jan 13, 2016·The Journal of Organic Chemistry·Rajendra Shivaji Mane, Bhalchandra M Bhanage
Oct 22, 2014·The Journal of Organic Chemistry·Nicole Nischan, Christian P R Hackenberger
Nov 20, 2009·Journal of Peptide Science : an Official Publication of the European Peptide Society·Dirk T S RijkersRob M J Liskamp
Apr 4, 2015·The Journal of Organic Chemistry·Sheng XieMingdi Yan
Mar 27, 2010·Chemistry & Biology·Emmanuel BasléMathieu Pucheault
Feb 20, 2014·The Journal of Organic Chemistry·Sachitanand M Mali, Hosahudya N Gopi
Sep 18, 2012·Organic Letters·Roberta CadoniLidia De Luca
Oct 29, 2015·Organic & Biomolecular Chemistry·Veladi PandurangaVommina V Sureshbabu
Apr 21, 2018·Organic & Biomolecular Chemistry·Narendra NVommina V Sureshbabu
Oct 30, 2010·Angewandte Chemie·Long YiRoger S Goody
May 29, 2012·Chembiochem : a European Journal of Chemical Biology·Ramiz NathaniStephen Caddick
Mar 29, 2014·Chemical Communications : Chem Comm·Michaela MühlbergChristian P R Hackenberger
Dec 22, 2009·Chemical Society Reviews·Luiz A CanalleJan C M van Hest
Sep 10, 2014·Chemical Communications : Chem Comm·Michael J GattnerMilan Vrabel
Oct 13, 2016·Chemistry : a European Journal·Kenward King Ho VongKatsunori Tanaka
Dec 14, 2018·Nature Reviews. Drug Discovery·Iriny EkladiousMark W Grinstaff
Dec 1, 2010·Chemical Communications : Chem Comm·Xiaohong ZhangChuan-Fa Liu
Jun 26, 2020·Chemistry : a European Journal·Ruairí O McCourt, Eoin M Scanlan
Feb 23, 2020·Organic Letters·Guillaume ClavéMichael Smietana
Aug 24, 2010·Organic Letters·Reda MhidiaOleg Melnyk
Jul 1, 2021·Journal of the American Chemical Society·Kota NomuraYasuhiro Kajihara

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