Exploiting Protein N-Terminus for Site-Specific Bioconjugation

Rosa, Lucía De; Stasi, Rossella Di; Romanelli, Alessandra; D’Andrea, Luca Domenico

doi:10.3390/molecules26123521

Cited by 25 publications

(27 citation statements)

References 116 publications

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“…The choice for a specific N-terminal targeting strategy will be highly application-dependent, and we refer the reader to recent reviews on N-terminal reactions for details. [77,78] The reaction may find use in affinity molecule decoration, to give for example ADCs. It will be of special interest to investigate combinations of azidogluconoylation with other conjugation strategies, apparently unaffected by gluconoylation, [43,79] but also to install more reactive handles onto the GDL-scaffold.…”

Section: Discussionmentioning

confidence: 99%

N‐Terminal Modification of Gly‐His‐Tagged Proteins with Azidogluconolactone

Brune

Lieknina

Sutov³

et al. 2021

ChemBioChem

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Site-specific protein modifications are vital for biopharmaceutical drug development. Gluconoylation is a nonenzymatic, post-translational modification of N-terminal HisTags. We report high-yield, site-selective in vitro α-aminoacylation of peptides, glycoproteins, antibodies, and Virus-like particles (VLPs) with azidogluconolactone at pH 7.5 in 1 h. Conjugates slowly hydrolyse, but diol-masking with borate esters inhibits reversibility. In an example, we multimerise azidogluconoylated SARS-CoV-2 receptor-binding domain (RBD) onto VLPs via click-chemistry, to give a COVID-19 vaccine. Compared to yeast antigen, HEK-derived RBD was immunologically superior, likely due to observed differences in glycosylation. We show the benefits of ordered over randomly oriented multimeric antigen display, by demonstrating single-shot seroconversion and best virus-neutralizing antibodies. Azidogluconoylation is simple, fast and robust chemistry, and should accelerate research and development.

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Section: Discussionmentioning

confidence: 99%

N‐Terminal Modification of Gly‐His‐Tagged Proteins with Azidogluconolactone

Brune

Lieknina

Sutov³

et al. 2021

ChemBioChem

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“…Under the non-specific modification, most proteins can lose their biological function and form oligomers or aggregate with further precipitation. Therefore, site-specific well-proved methods are required [79][80][81]. One of the possible proven procedures is the modification of only one cysteine No.…”

Section: Covalent Strategymentioning

confidence: 99%

Serum Albumin for Magnetic Nanoparticles Coating

Chubarov

2022

Magnetochemistry

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Magnetic nanoparticles (MNPs) have great potential in biochemistry and medical science. In particular, iron oxide nanoparticles have demonstrated a promising effect in various biomedical applications due to their high magnetic properties, large surface area, stability, and easy functionalization. However, colloidal stability, biocompatibility, and potential toxicity of MNPs in physiological environments are crucial for their in vivo application. In this context, many research articles focused on the possible procedures for MNPs coating to improve their physic-chemical and biological properties. This review highlights one viable fabrication strategy of biocompatible iron oxide nanoparticles using human serum albumin (HSA). HSA is mainly a transport protein with many functions in various fundamental processes. As it is one of the most abundant plasma proteins, not a single drug in the blood passes without its strength test. It influences the stability, pharmacokinetics, and biodistribution of different drug-delivery systems by binding or forming its protein corona on the surface. The development of albumin-based drug carriers is gaining increasing importance in the targeted delivery of cancer therapy. Considering this, HSA is a highly potential candidate for nanoparticles coating and theranostics area and can provide biocompatibility, prolonged blood circulation, and possibly resolve the drug-resistance cancer problem.

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“…Compared to the ubiquity of other reactive nucleophiles, N-terminal α-amine is appealing to site-selective chemical modification for being present in every protein and for consisting of a single reactive handle. Under the influence of the backbone amide, the α-amino group has a reduced pKa (6.0 to 8.0) compared to the ε-amine from lysines (~10.5), being highly reactive at near neutral conditions [85]. This difference in pKa can be exploited to achieve site-selective N-terminal labeling using amine modifying approaches, such as NHS-esters and reductive amination with aldehydes, under appropriate pH control [27,86,87].…”

Section: N-terminusmentioning

confidence: 99%

“…One of the issues concerning it, however, is developing chemoselective probes that can discriminate between the α-amino group and other nucleophiles, namely the other primary amines. As reviewed by De Rosa et al [85], several chemical strategies are already available for bioconjugation at the N-terminus. Thus, the remaining challenge is to expand the application of theses selective probes to the modifications of enzymes.…”

Section: Perspectivesmentioning

confidence: 99%

Protein Modifications: From Chemoselective Probes to Novel Biocatalysts

2021

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Chemical reactions can be performed to covalently modify specific residues in proteins. When applied to native enzymes, these chemical modifications can greatly expand the available set of building blocks for the development of biocatalysts. Nucleophilic canonical amino acid sidechains are the most readily accessible targets for such endeavors. A rich history of attempts to design enhanced or novel enzymes, from various protein scaffolds, has paved the way for a rapidly developing field with growing scientific, industrial, and biomedical applications. A major challenge is to devise reactions that are compatible with native proteins and can selectively modify specific residues. Cysteine, lysine, N-terminus, and carboxylate residues comprise the most widespread naturally occurring targets for enzyme modifications. In this review, chemical methods for selective modification of enzymes will be discussed, alongside with examples of reported applications. We aim to highlight the potential of such strategies to enhance enzyme function and create novel semisynthetic biocatalysts, as well as provide a perspective in a fast-evolving topic.

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Exploiting Protein N-Terminus for Site-Specific Bioconjugation

Cited by 25 publications

References 116 publications

N‐Terminal Modification of Gly‐His‐Tagged Proteins with Azidogluconolactone

N‐Terminal Modification of Gly‐His‐Tagged Proteins with Azidogluconolactone

Serum Albumin for Magnetic Nanoparticles Coating

Protein Modifications: From Chemoselective Probes to Novel Biocatalysts

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