Design of a synthetic luminescent probe from a biomolecule binding domain: selective detection of AU-rich mRNA sequences

Abstract

We report the design of a luminescent sensor based upon the zinc finger protein TIS11d, that allows for the selective time-resolved detection of the UUAUUUAUU sequence of the 3′-untranslated region of messenger RNA.

Using Native Chemical Ligation for Site‐Specific Synthesis of Hetero‐bis‐lanthanide Peptide Conjugates: Application to Ratiometric Visible or Near‐Infrared Detection of Zn2+

“…Design of hetero-bis-lanthanide peptide Zn 2 + + -responsive probes During the past years, we have described several responsive luminescent probes combining luminescent lanthanides, for their emission properties, and peptides, for their highly specific recognition properties. [30,[40][41][42][43] Especially,w eh aves hown that a classical bba zinc finger peptide with an appended DOTA-Ln 3 + Figure 1. Examples of emission spectrao fTb 3 + ,Eu 3 + ,Yb 3 + ,a nd Nd 3 + with DOTA-monoamide ligands obtained from compounds LZF1 Cs124 j Tb , LZF1 Cs124 j Eu ,and LZF1 NBD j Nd for Tb 3 + ,Eu 3 + ,and Nd 3 + , [30] respectively,and compound LZF3 Anthra j Yb for Yb 3 + (this work).…”

Luminescent Zinc Fingers: Zn‐Responsive Neodymium Near‐Infrared Emission in Water

“…Zn 2+ is involved in a large number of extracellular and intracellular signaling pathways and sensitive, selective, real‐time Zn 2+ ‐responsive probes are needed to understand and analyze these pathways . We are interested in creating responsive luminescent probes that combine Ln 3+ ions and peptide scaffolds, inspired by specific binding sites of proteins, to benefit from their unique recognition abilities . In order to design a Zn 2+ ‐responsive probe as a recognition unit, we opted for classical ββα zinc finger (ZF) peptides, which have a consensus sequence of the form (Tyr/Phe)‐Xaa‐Cys‐Xaa 2/4 ‐Cys‐Xaa 3 ‐Phe‐Xaa 5 ‐Leu‐Xaa 2 ‐His‐Xaa 3 ‐His, where Xaa can be any amino acid .…”

“…Ln 3 + complexesh ave desirable photophysical properties [12] for biological applicationsi ncluding:1)atom-like narrow emissionb ands at fixed wavelengthsi ndependent of experimental conditions and characteristico ft he nature of the Ln 3 + ion, 2) al arge difference betweent he absorption and emission wavelengths preventing self-absorption problemsa nd related quantification,3 )low or no tendencyt op hotobleaching, and 4) long emissionl ifetimes( micro to millisecond range), an advantage that can be used in time-gated detection experiments to suppress backgroundf luorescence of the biological medium (the lifetimes of which extend up to af ew nanoseconds for the longest). [22,23] In order to design a Zn 2 + -responsive probe as arecognition unit, we opted for classical bba zinc finger (ZF) peptides, which have ac onsensus sequenceo ft he form (Tyr/Phe)-Xaa-Cys-Xaa 2/4 -Cys-Xaa 3 -Phe-Xaa 5 -Leu-Xaa 2 -His-Xaa 3 -His, where Xaa can be any amino acid. [15][16][17][18][19] As main reasons, we can cite the limited number of antennae for the sensitization of NIR Ln 3 + luminescence and the efficient quenching of their emissionb yh armonicso fh igh energyO ÀHv ibrations of water molecules.…”

“…[21] We are interested in creatingr esponsive luminescent probest hat combine Ln 3 + ions andp eptides caffolds, inspired by specific binding sites of proteins,t ob enefit from their unique recognition abilities. [22,23] In order to design a Zn 2 + -responsive probe as arecognition unit, we opted for classical bba zinc finger (ZF) peptides, which have ac onsensus sequenceo ft he form (Tyr/Phe)-Xaa-Cys-Xaa 2/4 -Cys-Xaa 3 -Phe-Xaa 5 -Leu-Xaa 2 -His-Xaa 3 -His, where Xaa can be any amino acid. [24] They possess the advantage of selectively binding Zn 2 + among physiological cations [24,25] with their cysteine/histidine residues.W hen bound to Zn 2 + ,t hey adopt ac ompact bba fold (two b-sheets and an a-helix) due to the presence of three conserved amino acids (Tyr/Phe, Phe, and Leu) that constitute ah ydrophobic core within the structure.I nc ontrast, they are rather unfolded in the absence of Zn 2 + ( Figure 1A).…”

A novel DOTA-like building block with a picolinate arm for the synthesis of lanthanide complex-peptide conjugates with improved luminescence properties

“…Several peptide-and Ln 3+ -based responsive probes have been reported in the literature, in which the peptide acts as (i) a redox switch [33], (ii) a receptor for the targeted analyte, which can be a metal cation [39,50,51], a protein [52][53][54][55], an oligonucleotide [56][57][58] or (iii) the substrate of an enzyme of interest such as kinase or phosphatase [59][60][61]. The main interest of using a peptide as a recognition element in these probes is the selectivity it provides.…”