A doubly responsive probe for the detection of Cys4-tagged proteins

Kotera, Naoko; Dubost, Emmanuelle; Milanole, Gaëlle; Doris, Eric; Gravel, Edmond; Arhel, Nathalie J.; Dutasta, Jean‐Pierre; Cochrane, James R.; Mari, Emilie; Boutin, Céline; Léonce, Estelle; Berthault, Patrick; Rousseau, Bernard

doi:10.1039/c5cc04721h

Cited by 32 publications

(24 citation statements)

References 33 publications

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“…Most recently, Kotera et al attached hexa-carboxylate cryptophane to two arsenic moieties capable of interacting with proteins that contain a tetracysteine tag and observed a single peak that was shifted 6.4 ppm upon addition of excess Cys4-tagged peptide. 28 …”

Section: Cryptophane-based Xenon Biosensorsmentioning

confidence: 99%

An Expanded Palette of Xenon-129 NMR Biosensors

Wang

Dmochowski

2016

Acc. Chem. Res.

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ConspectusMolecular imaging holds considerable promise for elucidating biological processes in normal physiology as well as disease states, by determining the location and relative concentration of specific molecules of interest. Proton-based magnetic resonance imaging (1H MRI) is nonionizing and provides good spatial resolution for clinical imaging but lacks sensitivity for imaging low-abundance (i.e., submicromolar) molecular markers of disease or environments with low proton densities. To address these limitations, hyperpolarized (hp) 129Xe NMR spectroscopy and MRI have emerged as attractive complementary methodologies. Hyperpolarized xenon is nontoxic and can be readily delivered to patients via inhalation or injection, and improved xenon hyperpolarization technology makes it feasible to image the lungs and brain for clinical applications.In order to target hp 129Xe to biomolecular targets of interest, the concept of “xenon biosensing” was first proposed by a Berkeley team in 2001. The development of xenon biosensors has since focused on modifying organic host molecules (e.g., cryptophanes) via diverse conjugation chemistries and has brought about numerous sensing applications including the detection of peptides, proteins, oligonucleotides, metal ions, chemical modifications, and enzyme activity. Moreover, the large (∼300 ppm) chemical shift window for hp 129Xe bound to host molecules in water makes possible the simultaneous identification of multiple species in solution, that is, multiplexing. Beyond hyperpolarization, a 106-fold signal enhancement can be achieved through a technique known as hyperpolarized 129Xe chemical exchange saturation transfer (hyper-CEST), which shows great potential to meet the sensitivity requirement in many applications.This Account highlights an expanded palette of hyper-CEST biosensors, which now includes cryptophane and cucurbit[6]uril (CB[6]) small-molecule hosts, as well as genetically encoded gas vesicles and single proteins. In 2015, we reported picomolar detection of commercially available CB[6] via hyper-CEST. Inspired by the versatile host–guest chemistry of CB[6], our lab and others developed “turn-on” strategies for CB[6]-hyper-CEST biosensing, demonstrating detection of protein analytes in complex media and specific chemical events. CB[6] is starting to be employed for in vivo imaging applications. We also recently determined that TEM-1 β-lactamase can function as a single-protein reporter for hyper-CEST and observed useful saturation contrast for β-lactamase expressed in bacterial and mammalian cells. These newly developed small-molecule and genetically encoded xenon biosensors offer significant potential to extend the scope of hp 129Xe toward molecular MRI.

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Section: Cryptophane-based Xenon Biosensorsmentioning

confidence: 99%

An Expanded Palette of Xenon-129 NMR Biosensors

Wang

Dmochowski

2016

Acc. Chem. Res.

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“…Here, however, the concept is extended to the conception and use of a probe that is activatable in both fluorescence and 129 Xe NMR spectroscopy (i.e., the responses of these modalities change upon binding to the target). With respect to our previous study, the reported chemical shift modifications of caged xenon induced by binding of the probe to the target should be improved further for in cellulo biosensing applications. In this study, a set of four biosensors and three targeted peptide motifs were investigated to optimize the 129 Xe NMR response.…”

Section: Introductionmentioning

confidence: 81%

Bimodal Detection of Proteins by ¹²⁹Xe NMR and Fluorescence Spectroscopy

et al. 2019

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A full understanding of biological phenomena involves sensitive and noninvasive detection. Herein, we report the optimization of a probe for intracellular proteins that combines the advantages of fluorescence and hyperpolarized 129Xe NMR spectroscopy detection. The fluorescence detection part is composed of six residues containing a tetracysteine tag (−CCXXCC−) genetically incorporated into the protein of interest and of a small organic molecule, CrAsH. CrAsH becomes fluorescent if it binds to the tetracysteine tag. The part of the biosensor that enables detection by means of 129Xe NMR spectroscopy, which is linked to the CrAsH moiety by a spacer, is based on a cryptophane core that is fully suited to reversibly host xenon. Three different peptides, containing the tetracysteine tag and four organic biosensors of different stereochemistry, are benchmarked to propose the best couple that is fully suited for the in vitro detection of proteins.

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“…In cases of smaller targeting units (like short peptides), a dye like Cy3 can be directly integrated into the cryptophane conjugate [ 228 ] to achieve the same goal. This strategy was also pursued for a folate sensor [ 176 ], generation of bimodal probes for detecting proteins that are genetically incorporated with a tetracysteine tag [ 229 , 230 ] and the sensor for enabling cell surface glycan detection [ 128 , 231 ]. These are examples for validating a targeted cellular uptake/binding of functionalized Xe hosts via fluorophore-based detection.…”

Section: Aspects Of 129 Xe Biosensor Designmentioning

confidence: 99%

Molecular Sensing with Host Systems for Hyperpolarized 129Xe

Jayapaul

Schröder

2020

Molecules

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Hyperpolarized noble gases have been used early on in applications for sensitivity enhanced NMR. 129Xe has been explored for various applications because it can be used beyond the gas-driven examination of void spaces. Its solubility in aqueous solutions and its affinity for hydrophobic binding pockets allows “functionalization” through combination with host structures that bind one or multiple gas atoms. Moreover, the transient nature of gas binding in such hosts allows the combination with another signal enhancement technique, namely chemical exchange saturation transfer (CEST). Different systems have been investigated for implementing various types of so-called Xe biosensors where the gas binds to a targeted host to address molecular markers or to sense biophysical parameters. This review summarizes developments in biosensor design and synthesis for achieving molecular sensing with NMR at unprecedented sensitivity. Aspects regarding Xe exchange kinetics and chemical engineering of various classes of hosts for an efficient build-up of the CEST effect will also be discussed as well as the cavity design of host molecules to identify a pool of bound Xe. The concept is presented in the broader context of reporter design with insights from other modalities that are helpful for advancing the field of Xe biosensors.

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A doubly responsive probe for the detection of Cys4-tagged proteins

Cited by 32 publications

References 33 publications

An Expanded Palette of Xenon-129 NMR Biosensors

An Expanded Palette of Xenon-129 NMR Biosensors

Bimodal Detection of Proteins by ¹²⁹Xe NMR and Fluorescence Spectroscopy

Molecular Sensing with Host Systems for Hyperpolarized 129Xe

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A doubly responsive probe for the detection of Cys4-tagged proteins

Cited by 32 publications

References 33 publications

An Expanded Palette of Xenon-129 NMR Biosensors

An Expanded Palette of Xenon-129 NMR Biosensors

Bimodal Detection of Proteins by 129Xe NMR and Fluorescence Spectroscopy

Molecular Sensing with Host Systems for Hyperpolarized 129Xe

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Product

Resources

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Bimodal Detection of Proteins by ¹²⁹Xe NMR and Fluorescence Spectroscopy