Covalent protein–oligonucleotide conjugates by copper-free click reaction

Khatwani, Santoshkumar L.; Kang, Jun Sung; Mullen, Daniel G.; Hast, Michael A.; Beese, L.S.; Distefano, Mark D.; Taton, T. Andrew

doi:10.1016/j.bmc.2012.05.017

Cited by 39 publications

(38 citation statements)

References 65 publications

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“…More efficient results were obtained using bioorthogonal approaches in which the reactive groups do not modify the activity of the biomolecules. Specifically, bioorthogonal methods such as copper‐catalyzed azide‐alkyne [3+2] cycloaddition or oxime and hydrazone ligation have been proposed . In addition, several bifunctional cross‐linkers have been also described to produce protein conjugates ,.…”

Section: Methodsmentioning

confidence: 99%

Efficient bioactive oligonucleotide‐protein conjugation for cell‐targeted cancer therapy

et al. 2019

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Oligonucleotide‐protein conjugates have important applications in biomedicine. Simple and efficient methods are described for the preparation of these conjugates. Specifically, we describe a new method in which a bifunctional linker is attached to thiol‐oligonucleotide to generate a reactive intermediate that is used to link to the protein. Having similar conjugation efficacy compared with the classical method in which the bifunctional linker is attached first to the protein, this new approach produces significantly more active conjugates with higher batch to batch reproducibility. In a second approach, direct conjugation is proposed using oligonucleotides carrying carboxyl groups. These methodologies have been applied to prepare nanoconjugates of an engineered nanoparticle protein carrying a T22 peptide with affinity for the CXCR4 chemokine receptor and oligomers of the antiproliferative nucleotide 2’‐deoxy‐5‐fluorouridine in a very efficient way. The protocols have potential uses for the functionalization of proteins, amino‐containing polymers or amino‐lipids in order to produce complex therapeutic nucleic acid delivery systems.

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Section: Methodsmentioning

confidence: 99%

Efficient bioactive oligonucleotide‐protein conjugation for cell‐targeted cancer therapy

et al. 2019

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“…Aiming to address this limitation, Dozier et al analysed how different FTase mutations within the active site could be used to better accommodate these larger analogues. 259 Based on the crystal structures of FTase and previously reported literature, 260 three key residues were identified that were in close contact with the third isoprenoid unit of FPP. Accordingly, three mutants were prepared, including W102A, Y154A, and Y205A.…”

Section: Enzymatic Protein Labelling Strategiesmentioning

confidence: 99%

Recent progress in enzymatic protein labelling techniques and their applications

et al. 2018

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Protein-based conjugates are valuable constructs for a variety of applications. Conjugation of proteins to fluorophores is commonly used to study their cellular localization and the protein-protein interactions. Modification of therapeutic proteins with either polymers or cytotoxic moieties greatly enhances their pharmacokinetics and potency. To label a protein of interest, conventional direct chemical reaction with the side-chains of native amino acids often yields heterogeneously modified products. This renders their characterization complicated, requires difficult separation steps and may impact protein function. Although modification can also be achieved via the insertion of unnatural amino acids bearing bioorthogonal functional groups, these methods can have lower protein expression yields, limiting large scale production. As a site-specific modification method, enzymatic protein labelling is highly efficient and robust under mild reaction conditions. Significant progress has been made over the last five years in modifying proteins using enzymatic methods for numerous applications, including the creation of clinically relevant conjugates with polymers, cytotoxins or imaging agents, fluorescent or affinity probes to study complex protein interaction networks, and protein-linked materials for biosensing. This review summarizes developments in enzymatic protein labelling over the last five years for a panel of ten enzymes, including sortase A, subtiligase, microbial transglutaminase, farnesyltransferase, N-myristoyltransferase, phosphopantetheinyl transferases, tubulin tyrosin ligase, lipoic acid ligase, biotin ligase and formylglycine generating enzyme.

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“…To reduce the toxic damage of copper ions, Khatwani et al used a copperfree click reaction (e.g. strain-promoted [3 + 2] azide-alkyne cycloaddition) to produce protein-DNA conjugates [51]. Francis and coworkers reported a protein bioconjugation at an N-terminal amino acid via a pyridoxal 5′-phosphate (PLP)mediated transamination reaction [52], and Takeda et al reported the development of a method to chemically modify an oligonucleotide to a N-terminal cysteine of a protein [53].…”

Section: Protein-dna Bioconjugationmentioning

confidence: 99%

DNA-Scaffolded Proximity Assembly and Confinement of Multienzyme Reactions

Wang

Liang

et al. 2020

Top Curr Chem (Z)

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Cellular functions rely on a series of organized and regulated multienzyme cascade reactions. The catalytic efficiencies of these cascades depend on the precise spatial organization of the constituent enzymes, which is optimized to facilitate substrate transport and regulate activities. Mimicry of this organization in a non-living, artificial system would be very useful in a broad range of applications-with impacts on both the scientific community and society at large. Self-assembled DNA nanostructures are promising applications to organize biomolecular components into prescribed, multidimensional patterns. In this review, we focus on recent progress in the field of DNA-scaffolded assembly and confinement of multienzyme reactions. DNA self-assembly is exploited to build spatially organized multienzyme cascades with control over their relative distance, substrate diffusion paths, compartmentalization and activity actuation. The combination of addressable DNA assembly and multienzyme cascades can deliver breakthroughs toward the engineering of novel synthetic and biomimetic reactors.

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Covalent protein–oligonucleotide conjugates by copper-free click reaction

Cited by 39 publications

References 65 publications

Efficient bioactive oligonucleotide‐protein conjugation for cell‐targeted cancer therapy

Efficient bioactive oligonucleotide‐protein conjugation for cell‐targeted cancer therapy

Recent progress in enzymatic protein labelling techniques and their applications

DNA-Scaffolded Proximity Assembly and Confinement of Multienzyme Reactions

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