Lanthanide-based peptide biosensor to monitor CDK4/cyclin D kinase activity

González‐Vera, Juan A.; Bouzada, David; Bouclier, Céline; Vázquez, M. Eugenio; Morris, May

doi:10.1039/c6cc09948c

Cited by 20 publications

(18 citation statements)

References 23 publications

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“…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.…”

Section: Introductionmentioning

confidence: 99%

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

Fremy

Raibaut

Cepeda

et al. 2020

Journal of Inorganic Biochemistry

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Combination of complexes of trivalent lanthanide cations (Ln 3+) for their luminescent properties and peptides for their recognition properties or folding abilities is interesting in view of designing responsive luminescent probes. The octadentate DOTA chelate is the most popular chelate to design luminescent Ln 3+ complex-peptide conjugates. In this article, we describe a novel building block, DO3Apic-tris(allyl)ester, which provides access to peptides with a conjugated nonadentate chelate, namely DO3Apic, featuring a cyclen macrocycle functionalized by three acetate and one picolinamide arms, for improved luminescence properties. This building block, with allyl protecting groups, is readily obtained by solid phase synthesis. We show that it is superior to its analogue with tBu protecting groups for the preparation of peptide conjugates because of the difficult removal of the tBu protecting groups for the latter. Then, two Zn 2+-responsive luminescent probes, which rely on (i) a zinc finger scaffold for selective Zn 2+ binding, (ii) a Eu 3+ complex and (iii) an acridone antenna for long-wavelength sensitization of Eu 3+ luminescence, are compared. One of these probes, LZF3 ACD|Eu , incorporates a DOTA chelate whereas the other, LZF4 ACD|Eu , incorporates a DO3Apic chelate. We show that changing the octadentate DOTA for the nonadentate DO3Apic ligand results in a higher Eu 3+ luminescence lifetime and in a doubling of the quantum yield, confirming the interest of the DO3Apic chelate and the DO3Apic(tris(allyl)ester building block for the preparation of Ln 3+ complex-peptide conjugates. Additionally, the DO3Apic chelate provides self-calibration for LZF4 ACD|Eu luminescence upon excitation of its picolinamide chromophore, making LZF4 ACD|Eu a ratiometric sensor for Zn 2+ detection.

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Section: Introductionmentioning

confidence: 99%

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

Fremy

Raibaut

Cepeda

et al. 2020

Journal of Inorganic Biochemistry

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“…Please do not adjust margins Conversely, the alkaline phosphatase catalysed dephosphorylation reaction could be monitored by observing an increase in Eu(III) emission intensity upon preferential binding to the β-diketonate antenna. More recently, an assay for CDK4/cyclin D kinase activity was developed (Figure 6c), 119 capable of operating in melanoma cell extracts and reporting on CDK4 inhibition. The Tb(III)-based biosensor comprises a CDK4 substrate sequence attached to a recognition domain for the phosphorylated peptide.…”

Section: Please Do Not Adjust Marginsmentioning

confidence: 99%

Application of lanthanide luminescence in probing enzyme activity

Hewitt

Butler

2018

Chem. Commun.

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Enzymes play critical roles in the regulation of cellular function and are implicated in numerous disease conditions. Reliable and practicable assays are required to study enzyme activity, to facilitate the discovery of inhibitors and activators of enzymes related to disease. In recent years, a variety of enzyme assays have been devised that utilise luminescent lanthanide(iii) complexes, taking advantage of their high detection sensitivities, long luminescence lifetimes, and line-like emission spectra that permit ratiometric and time-resolved analyses. In this Feature article, we focus on recent progress in the development of enzyme activity assays based on lanthanide(iii) luminescence, covering a variety of strategies including Ln(iii)-labelled antibodies and proteins, Ln(iii) ion encapsulation within defined peptide sequences, reactivity-based Ln(iii) probes, and discrete Ln(iii) complexes. Emerging approaches for monitoring enzyme activity are discussed, including the use of anion responsive lanthanide(iii) complexes, capable of molecular recognition and luminescence signalling of polyphosphate anions.

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“…Fluorescent peptide/peptidomimetic-based targeting and visualization of the subcellular localization, 1,2 and probing and modulation of intra/inter-biomolecular interactions, 3,4 of the protein target of interest provides a versatile platform, either in vitro/vivo or in assays, [5][6][7] for fundamental biochemistry/ chemical biology research, [8][9][10] as well as drug discovery and development. [11][12][13][14] With the growing signicance and recent successes in peptide (and peptide-containing) therapeutics, [15][16][17][18][19][20] by virtue of their (i) scalable modular synthetic accessibility and tunability, (ii) target-specic selectivity, potency and efficacy, (iii) good biocompatibility, low systemic toxicity and easy clearance, there is a great demand for more general and exible synthetic methodologies and ligation protocols to make peptides uorescent, upon functional de novo design, for indepth biomedical investigations.…”

Section: Introductionmentioning

confidence: 99%