2017
DOI: 10.1039/c6cc09948c
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Lanthanide-based peptide biosensor to monitor CDK4/cyclin D kinase activity

Abstract: We describe a lanthanide biosensor that responds to CDK4 kinase activity in melanoma cell extracts through a significant and dose dependent increase in luminescence, thanks to sensitization of a DOTA[Tb] complex incorporated into a CDK4 substrate peptide by a unique tryptophan residue in an adjacent phosphoaminoacid binding moiety.

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Cited by 21 publications
(18 citation statements)
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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%
“…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%
“…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%
“…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%