Click conjugation of a binuclear terbium(III) complex for real-time detection of tyrosine phosphorylation

Analytical Chemistry
Hiroki AkibaMakoto Komiyama

Abstract

Phosphorylation of proteins is closely associated with various diseases, and, therefore, its detection is vitally important in molecular biology and drug discovery. Previously, we developed a binuclear Tb(III) complex, which emits notable luminescence only in the presence of phosphotyrosine. In this study, we conjugated a newly synthesized binuclear Tb(III) complex to substrate peptides by using click chemistry. Using these conjugates, we were able to detect tyrosine phosphorylation in real time. These conjugates were superior to nonconjugated Tb(III) complexes for the detection of tyrosine phosphorylation, especially when the substrate peptides used were positively charged. Luminescence intensity upon phosphorylation was enhanced 10-fold, making the luminescence intensity of this system one of the largest among lanthanide luminescence-based systems. We also determined Michaelis-Menten parameters for the phosphorylation of various kinase/peptide combinations and quantitatively analyzed the effects of mutations in the peptide substrates. Furthermore, we successfully monitored the inhibition of enzymatic phosphorylation by inhibitors in real time. Advantageously, this system detects only the phosphorylation of tyrosine (phosphory...Continue Reading

References

May 25, 1992·Journal of Inorganic Biochemistry·S L Klakamp, W D Horrocks
Jan 1, 1982·Proceedings of the National Academy of Sciences of the United States of America·J E CasnellieE G Krebs
Feb 7, 2004·Chemical Communications : Chem Comm·Paul AtkinsonDavid Parker
Jul 7, 2005·Journal of the American Chemical Society·Lucy L Liu, Katherine J Franz
Dec 12, 2007·Organic Letters·Matthew S TremblayDalibor Sames
Feb 16, 2010·Chemical Reviews·Jean-Claude G Bünzli
Apr 7, 2010·Journal of the American Chemical Society·Susan Carr ZondloNeal J Zondlo
Apr 15, 2010·Chemistry : a European Journal·Hiroki AkibaMakoto Komiyama
Oct 7, 2011·Chemical Society Reviews·Juan A González-Vera
Nov 1, 2011·Nature Biotechnology·Theonie AnastassiadisJeffrey R Peterson
Nov 1, 2011·Nature Biotechnology·Mindy I DavisPatrick P Zarrinkar
Dec 14, 2011·Chemistry : a European Journal·Candy H S LuShao Q Yao
Jul 5, 2012·Chemical Society Reviews·Stephen J Butler, David Parker
Aug 18, 2012·Chemical Communications : Chem Comm·Elena PazosM Eugenio Vázquez
Feb 19, 2013·Analytical Chemistry·Andrew M Lipchik, Laurie L Parker
Dec 21, 2013·Biotechnology Journal·Camille PrévelMay C Morris

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Citations

Jul 5, 2016·International Journal of Analytical Chemistry·Jun SumaokaMakoto Komiyama
May 24, 2018·Chemical Communications : Chem Comm·Sarah H Hewitt, Stephen J Butler
Apr 29, 2018·Scientific Reports·Jessica ShakleeJop H van Berlo
Sep 6, 2018·Journal of Medicinal Chemistry·Mingcheng QianSerge Van Calenbergh
Sep 8, 2020·Journal of the American Chemical Society·Minmin LiGuangyan Qing

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