Transglutaminase and Sialyltransferase Enzymatic Approaches for Polymer Conjugation to Proteins

Maso, Katia; Grigoletto, Antonella; Pasut, Gianfranco

doi:10.1016/bs.apcsb.2018.01.003

Cited by 9 publications

(7 citation statements)

References 68 publications

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“…Chilkoti et al demonstrated that C-terminal modification of exendin-4 using sortase A yielded conjugates that were able to reduce blood glucose levels for longer than the FDA-approved PEGylated drugs without causing an immune response . Transglutaminase and sialyltransferase have also been used as ligation mediators . Transglutaminase uses an acyltransferase mechanism to catalyze the transfer of the acyl group of a γ-carboxamide (acyl donor) of a glutamine residue to the γ-amine (acyl acceptor) of a lysine residue.…”

Section: Protein–polymer Conjugatesmentioning

confidence: 99%

“…Transglutaminase uses an acyltransferase mechanism to catalyze the transfer of the acyl group of a γ-carboxamide (acyl donor) of a glutamine residue to the γ-amine (acyl acceptor) of a lysine residue. Therefore, either the protein amino acid to be modified is a Gln with a polymer containing a primary amine, or the amino acid is a Lys with a polymer containing a Gln moiety . Additionally, sialyltransferases are involved in protein glycosylation and have been used to site-specifically PEGylate a protein, and the process was deemed GlycoPEGylation.…”

Section: Protein–polymer Conjugatesmentioning

confidence: 99%

“…124 Transglutaminase and sialyltransferase have also been used as ligation mediators. 125 Transglutaminase uses an acyltransferase mechanism to catalyze the transfer of the acyl group of a γ-carboxamide (acyl donor) of a glutamine residue to the γ-amine (acyl acceptor) of a lysine residue. Therefore, either the protein amino acid to be modified is a Gln with a polymer containing a primary amine, or the amino acid is a Lys with a polymer containing a Gln moiety.…”

Section: ■ Protein−polymer Conjugatesmentioning

confidence: 99%

“…Therefore, either the protein amino acid to be modified is a Gln with a polymer containing a primary amine, or the amino acid is a Lys with a polymer containing a Gln moiety. 125 Additionally, sialyltransferases are involved in protein glycosylation and have been used to sitespecifically PEGylate a protein, and the process was deemed GlycoPEGylation. A PEGylated nucleotide sugar was covalently conjugated to the carbohydrate group of a protein.…”

Section: ■ Protein−polymer Conjugatesmentioning

confidence: 99%

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Atom Transfer Radical Polymerization for Biorelated Hybrid Materials

et al. 2019

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Proteins, nucleic acids, lipid vesicles, and carbohydrates are the major classes of biomacromolecules that function to sustain life. Biology also uses post-translation modification to increase the diversity and functionality of these materials, which has inspired attaching various other types of polymers to biomacromolecules. These polymers can be naturally (carbohydrates and biomimetic polymers) or synthetically derived and have unique properties with tunable architectures. Polymers are either grafted-to or grown-from the biomacromolecule's surface, and characteristics including polymer molar mass, grafting density, and degree of branching can be controlled by changing reaction stoichiometries. The resultant conjugated products display a chimerism of properties such as polymer-induced enhancement in stability with maintained bioactivity, and while polymers are most often conjugated to proteins, they are starting to be attached to nucleic acids and lipid membranes (cells) as well. The fundamental studies with protein−polymer conjugates have improved our synthetic approaches, characterization techniques, and understanding of structure−function relationships that will lay the groundwork for creating new conjugated biomacromolecular products which could lead to breakthroughs in genetic and tissue engineering.

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Section: Protein–polymer Conjugatesmentioning

confidence: 99%

Section: Protein–polymer Conjugatesmentioning

confidence: 99%

Section: ■ Protein−polymer Conjugatesmentioning

confidence: 99%

Section: ■ Protein−polymer Conjugatesmentioning

confidence: 99%

See 2 more Smart Citations

Atom Transfer Radical Polymerization for Biorelated Hybrid Materials

et al. 2019

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“…Therefore, it is possible to achieve site-selective modification on the endogenous glutamine residues without genetic engineering. Such examples included the PEGylation of human growth factor (hGH, Q40 and Q141), 220 human interleukin-2 (hIL-2, Q74), 221 granulocyte colony stimulating factor (GSCF, Q134), 222 and more recently interferon alpha-2b (IFN α−2b, Q101). 223 To further increase the selectivity of MTG, Pasut and coworkers explored the use of organic co-solvents, which was known to influence protein conformation and flexibility.…”

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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Chemoselective Methionine Labelling of Recombinant Trastuzumab Shows High In Vitro and In Vivo Tumour Targeting

Hashad

Jap

Casey

et al. 2023

Chemistry A European J

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A highly effective 2‐step system for site‐specific antibody modification and conjugation of the monoclonal antibody Herceptin (commercially available under Trastuzumab) in a cysteine‐independent manner was used to generate labelled antibodies for in vivo imaging. The first step contains redox‐activated chemical tagging (ReACT) of thioethers via engineered methionine residues to introduce specific alkyne moieties, thereby offering a novel easy way to fundamentally change the process of antibody bioconjugation. The second step involves modification of the introduced alkyne via azide‐alkyne cycloaddition ′click′ conjugation. The versatility of this 2‐step approach is demonstrated here by the selective incorporation of a fluorescent dye but can also be applied to a wide variety of different conjugation partners depending on the desired application in a facile manner. Methionine‐modified antibodies were characterised in vitro, and the diagnostic potential of the most promising variant was further analysed in an in vivo xenograft animal model using a fluorescence imaging modality. This study demonstrates how methionine‐mediated antibody conjugation offers an orthogonal and versatile route to the generation of tailored antibody conjugates with in vivo applicability.

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Transglutaminase and Sialyltransferase Enzymatic Approaches for Polymer Conjugation to Proteins

Cited by 9 publications

References 68 publications

Atom Transfer Radical Polymerization for Biorelated Hybrid Materials

Atom Transfer Radical Polymerization for Biorelated Hybrid Materials

Recent progress in enzymatic protein labelling techniques and their applications

Chemoselective Methionine Labelling of Recombinant Trastuzumab Shows High In Vitro and In Vivo Tumour Targeting

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