Peptide‐Directed DNA‐Templated Protein Labelling for The Assembly of a Pseudo‐IgM

Nielsen, Thorbjørn B.; Thomsen, Rasmus P.; Mortensen, Michael; Kjems, Jørgen; Nielsen, Per F.; Kodal, Anne Louise Bank; Cló, Emiliano; Gothelf, Kurt V.

doi:10.1002/anie.201903134

“…DPAL has been used to identify the target of small molecules,p eptides, nucleic acids,a nd aptamers. [13,14] With as imilar concept, the Gothelf group developed an elegant method for site-specific antibody-DNAc onjugation, [15] and the Tang roup used aptamers to guide DNA-templated protein conjugation. [16] However,D PA Lh as not been exploited to study proteinprotein interactions (PPIs).…”

Section: Resultsmentioning

confidence: 99%

Identification of Histone deacetylase (HDAC)‐Associated Proteins with DNA‐Programmed Affinity Labeling

Zhang

¹

,

Peng

²

,

Huang

³

et al. 2020

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Histone deacetylase (HDAC) is a major class of deacetylation enzymes. Many HDACs exist in large protein complexes in cells and their functions strongly depend on the complex composition. The identification of HDAC‐associated proteins is highly important in understanding their molecular mechanisms. Although affinity probes have been developed to study HDACs, they were mostly targeting the direct binder HDAC, while other proteins in the complex remain underexplored. We report a DNA‐based affinity labeling method capable of presenting different probe configurations without the need for preparing multiple probes. Using one binding probe, 9 probe configurations were created to profile HDAC complexes. Notably, this method identified indirect HDAC binders that may be inaccessible to traditional affinity probes, and it also revealed new biological implications for HDAC‐associated proteins. This study provided a simple and broadly applicable method for characterizing protein‐protein interactions.

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“…DPAL has been used to identify the target of small molecules,p eptides, nucleic acids,a nd aptamers. [13,14] With as imilar concept, the Gothelf group developed an elegant method for site-specific antibody-DNAc onjugation, [15] and the Tang roup used aptamers to guide DNA-templated protein conjugation. [16] However,D PA Lh as not been exploited to study proteinprotein interactions (PPIs).…”

Section: Resultsmentioning

confidence: 99%

Identification of Histone deacetylase (HDAC)‐Associated Proteins with DNA‐Programmed Affinity Labeling

Zhang

¹

,

Peng

²

,

Huang

³

et al. 2020

Angewandte Chemie

3

0

View full text Add to dashboard Cite

Histone deacetylase (HDAC) is a major class of deacetylation enzymes. Many HDACs exist in large protein complexes in cells and their functions strongly depend on the complex composition. The identification of HDAC‐associated proteins is highly important in understanding their molecular mechanisms. Although affinity probes have been developed to study HDACs, they were mostly targeting the direct binder HDAC, while other proteins in the complex remain underexplored. We report a DNA‐based affinity labeling method capable of presenting different probe configurations without the need for preparing multiple probes. Using one binding probe, 9 probe configurations were created to profile HDAC complexes. Notably, this method identified indirect HDAC binders that may be inaccessible to traditional affinity probes, and it also revealed new biological implications for HDAC‐associated proteins. This study provided a simple and broadly applicable method for characterizing protein‐protein interactions.

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“…Authors of this study used these DNA-conjugated antibodies to assemble IgG pentamers around a core DNA motif, mimicking the natural IgM structure. 71 …”

Section: Affinity Ligands and Their Role In Site-specific Conjugation Of Unmodified Antibodiesmentioning

confidence: 99%

Affinity-Based Methods for Site-Specific Conjugation of Antibodies

Witting

¹

,

Hober

²

,

Kanje

³

2021

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Conjugation of various reagents to antibodies has long been an elegant way to combine the superior binding features of the antibody with other desired but non-natural functions. Applications range from labels for detection in different analytical assays to the creation of new drugs by conjugation to molecules which improves the pharmaceutical effect. In many of these applications, it has been proven advantageous to control both the site and the stoichiometry of the conjugation to achieve a homogeneous product with predictable, and often also improved, characteristics. For this purpose, many research groups have, during the latest decade, reported novel methods and techniques, based on small molecules, peptides, and proteins with inherent affinity for the antibody, for site-specific conjugation of antibodies. This review provides a comprehensive overview of these methods and their applications and also describes a historical perspective of the field.

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Peptide‐Directed DNA‐Templated Protein Labelling for The Assembly of a Pseudo‐IgM

Cited by 32 publications

References 31 publications

Identification of Histone deacetylase (HDAC)‐Associated Proteins with DNA‐Programmed Affinity Labeling

Identification of Histone deacetylase (HDAC)‐Associated Proteins with DNA‐Programmed Affinity Labeling

Identification of Histone deacetylase (HDAC)‐Associated Proteins with DNA‐Programmed Affinity Labeling

Affinity-Based Methods for Site-Specific Conjugation of Antibodies

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