Tyrosine bioconjugation – an emergent alternative

Szijj, Péter A.; Kostadinova, Kristina A; Spears, Richard J.; Chudasama, Vijay

doi:10.1039/d0ob01912g

Cited by 57 publications

(66 citation statements)

References 60 publications

(108 reference statements)

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“…Promising approaches have recently been described to overcome the initial limitations of Lys 10,11 and Cys 12,13 modifications, and much effort is now dedicated to selectively target less exploited amino acids such as methionine, 14 tryptophan, 15,16 histidine 17,18 or tyrosine (Y). 19,20 Y usually occurs at low frequency in proteins (<5%) and is partially buried in the surface, offering unique opportunities for controlled labelling of the most reactive residues. 21,22 The phenol side chain of Y is neutral at physiological pH and experiences several biologically relevant post-translational modifications (phosphorylations, sulfations, nitrations, Oglycosylations, oxydations).…”

Section: Introductionmentioning

confidence: 99%

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Luminol anchors improve the electrochemical-tyrosine-click labelling of proteins

et al. 2021

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Luminol anchors improve the electrochemical-tyrosine-click labelling of proteins

et al. 2021

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“…Promising approaches have been recently described to overcome the initial limitations of Lys 10,11 and Cys 12,13 modifications, and much efforts are now dedicated to selectively target less exploited amino acids such as methionine, 14 tryptophan, 15,16 histidine 17,18 or tyrosine (Y). 19,20 Y usually occur with low frequency in proteins (~3-4%) and are partially buried at the surface, offering unique opportunities for a controlled labelling of the most reactive residues. 21 The phenol side chain of Y is neutral at physiological pH and experience several post-translational modifications of biological relevance (phosphorylations, sulfations, nitrations, Oglycosylations, oxydations).…”

Section: Introductionmentioning

confidence: 99%

Luminol Anchors Improve the Electrochemical-Tyrosine-Click Labelling of Proteins

Depienne

Álvarez-Dorta

Croyal

et al. 2021

Preprint

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New methodologies for the chemo-selective modifications of peptides and native proteins are of great importance in chemical biology and for the developm ent of therapeutic conjugates. Less abundant and uncharged amino-acid residues are interesting targets to form less heterogeneous conjugates and preserve biological functions. Phenylurazole (PhUr), Nmethylphenylurazole (NMePhUr) and N-methylluminol (NMeLum) derivatives were described as tyrosine (Y) anchors after chemical or enzymatic oxydations. Recently, we developed the first electrochemical Y-bioconjugation method coined eYclick to activate PhUr in biocompatible media. In this work, we assessed the limitations, benefits and relative efficiencies of eY-click conjugations performed with a set of PhUr, NMePhUr and NMeLum derivatives. Results evidenced a high efficiency of NMeLum that showed a complete Y-chemoselectivity on polypeptides and biologically relevant proteins after soft electrochemical activation. Side reactions on nucleophilic or heteroaromatic amino-acids such as lysine or tryptophan were never observed during mass spectrometry analysis. Myoglobine, bovine serum albumin, a plant mannosidase, glucose oxidase and the therapeutically relevant antibody trastuzumab were efficiently labelled with a fluorescent probe in a twostep approach combining eY-click and strain-promoted azide-alkyne cyclization (SPAAC). The proteins conserved their structural integrity as observed by circular dichroism and the trastuzumab conjugate showed a similar binding affinity for the natural HER2 ligand as shown by bio-layer interferometry. Compared to our previously described protocol with PhUr, eY-click with NMeLum species showed faster reaction kinetics, higher (complete) Y-chemoselectivity and reactivity, and offer the interesting possibility for the double tagging of solvent-exposed Y.

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“…Building on the foundations of Pauly, [7] Inoue, and Landsteiner [8] as detailed by Higgins et al, [9.10] tyrosine (Tyr/Y) residues have piqued the interests of the bioconjugation community [11] . For excellent reviews of tyrosine conjugation methods for labelling of antibodies and proteins see Szijj et al [12] . and Dorta et al [13] .…”

Section: Introductionmentioning

confidence: 99%

Bio‐Orthogonal Conjugation of a Cationic Metalloporphyrin to BSA and HSA via “Click” Chemistry

et al. 2021

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In this study, we present a convenient method for the labelling of tyrosine residues on bovine serum albumin (BSA) and human serum albumin (HSA) and report for the first time their subsequent bio‐orthogonal conjugation with porphyrins via “click” chemistry. We demonstrate that these serum proteins can be labelled with an alkyne‐diazonium heterobifunctional linker and can then undergo chemo‐selective bio‐orthogonal conjugation with a water‐soluble azido metalloporphyrin via “click” chemistry to yield protein‐conjugates that retain their photodynamic properties. In our hands, this method was found to be highly reproducible, scalable, and tuneable which allows for the production of bioconjugates where the porphyrin‐protein conjugate not only retains an ability to generate singlet oxygen but possess an enhanced relative singlet oxygen quantum yields relative to the porphyrin alone. Furthermore, we have investigated the photochemical properties of these conjugates through photospectrometric techniques and have determined that the porphyrin macrocycles remain appreciably photostable under light irradiation. Our phototoxic protein‐photosensitizer‐conjugates show excellent photodynamic activity against a human colorectal adenocarcinoma cancer cell line (HT‐29) with cell viabilities of 7.7±0.5 % (IC50 8.76±2.14 μM) and 1.7±1.9 % (IC50 8.48±5.11 μM) for BSA and HAS, respectively, when irradiated with 20 J cm−2 of white‐light. Importantly, neither of the conjugates was found to possess any significant “dark” toxicity even at concentrations of 100 μM. Furthermore, the natural fluorescent properties of the bioconjugates allowed for the determination of cellular uptake in vitro via fluorescence microscopy thus highlighting the potential theranostic applications of these unique protein‐drug‐conjugates.

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Tyrosine bioconjugation – an emergent alternative

Abstract: Protein bioconjugation is an increasingly important field of research, with wide-ranging applications in areas such as therapeutics and biomaterials. Traditional cysteine and lysine bioconjugation strategies are widely used and have...

Cited by 57 publications

References 60 publications

Luminol anchors improve the electrochemical-tyrosine-click labelling of proteins

Luminol anchors improve the electrochemical-tyrosine-click labelling of proteins

Luminol Anchors Improve the Electrochemical-Tyrosine-Click Labelling of Proteins

Bio‐Orthogonal Conjugation of a Cationic Metalloporphyrin to BSA and HSA via “Click” Chemistry

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