Selective Sensing of Tyrosine Phosphorylation in Peptides Using Terbium(III) Complexes

Abstract

Phosphorylation of tyrosine residues in proteins, as well as their dephosphorylation, is closely related to various diseases. However, this phosphorylation is usually accompanied by more abundant phosphorylation of serine and threonine residues in the proteins and covers only 0.05% of the total phosphorylation. Accordingly, highly selective detection of phosphorylated tyrosine in proteins is an urgent subject. In this review, recent developments in this field are described. Monomeric and binuclear TbIII complexes, which emit notable luminescence only in the presence of phosphotyrosine (pTyr), have been developed. There, the benzene ring of pTyr functions as an antenna and transfers its photoexcitation energy to the TbIII ion as the emission center. Even in the coexistence of phosphoserine (pSer) and phosphothreonine (pThr), pTyr can be efficintly detected with high selectivity. Simply by adding these TbIII complexes to the solutions, phosphorylation of tyrosine in peptides by protein tyrosine kinases and dephosphorylation by protein tyrosine phosphatases can be successfully visualized in a real-time fashion. Furthermore, the activities of various inhibitors on these enzymes are quantitatively evaluated, indicating a strong potential of the method for efficient screening of eminent inhibitors from a number of candidates.

Methods for the recombinant expression of active tyrosine kinase domains: Guidelines and pitfalls

“…LanthaScreen assays measure the fluorescence energy transfer (FRET) from a phospho-tyrosine antibody labelled with a donor fluorophore to the acceptor fluorescent dye that is conjugated to a substrate peptide. The method uses the exceptionally long fluorescence life time of lanthanide ions (europium or terbium), which allows to delay the measurement of donor and acceptor emission until background fluorescence from other potentially interfering compounds has decayed (Sumaoka, Akiba, & Komiyama, 2016;Carlson, Robers, Vogel, & Machleidt, 2009).…”

Recent Advances in Protein Kinase Activity Analysis Based on Nanomaterials

“…Recently, Sumaoka and co-workers demonstrated that lanthanide ions and their complexes such as Terbium (III) complexes show high selectivity to phosphotyrosine (pTyr) [42]. Another similar study was reported by Hussain et al, who fabricated diamond-lanthanum oxide and diamond-samarium oxide nanocomposites and applied them to selectively capture phosphorylated peptides from β-casein and BSA.…”

In vitro methods for testing antiviral drugs

“…Kinase-catalyzed phosphorylation of fluorescent peptides promotes their binding to metal-coated nanoparticles, which decreases their mobility and enhances measurable fluorescence polarization (Lea and Simeonov, 2011;Sportsman et al, 2004). TbIII complexes, in which phosphotyrosine induces fluorescence emission (Wang et al, 2015a), may be used to evaluate protein tyrosine kinase activity (Akiba et al, 2015;Sumaoka et al, 2016). Fluorescence polarization methods also can be useful for drug screening [reviewed in (Hall et al, 2016)].…”