Kinetic Controlled Tag-Catcher Interactions for Directed Covalent Protein Assembly

Tan, Lee Ling; Hoon, Shawn; Wong, Fong Tian

doi:10.1371/journal.pone.0165074

Cited by 54 publications

(47 citation statements)

References 28 publications

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“…To address whether the C-terminal residues are dispensable for binding or do even negatively affect the interaction as observed for SdyTag [35], a C-terminal truncation of the tag by three residues (R257-N259) was proposed (4oq1 T (ΔRGN)).…”

Section: Resultsmentioning

confidence: 99%

Probing the potential of CnaB-type domains for the design of tag/catcher systems

et al. 2017

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Building proteins into larger, post-translational assemblies in a defined and stable way is still a challenging task. A promising approach relies on so-called tag/catcher systems that are fused to the proteins of interest and allow a durable linkage via covalent intermolecular bonds. Tags and catchers are generated by splitting protein domains that contain intramolecular isopeptide or ester bonds that form autocatalytically under physiological conditions. There are already numerous biotechnological and medical applications that demonstrate the usefulness of covalent linkages mediated by these systems. Additional covalent tag/catcher systems would allow creating more complex and ultra-stable protein architectures and networks. Two of the presently available tag/catcher systems were derived from closely related CnaB-domains of Streptococcus pyogenes and Streptococcus dysgalactiae proteins. However, it is unclear whether domain splitting is generally tolerated within the CnaB-family or only by a small subset of these domains. To address this point, we have selected a set of four CnaB domains of low sequence similarity and characterized the resulting tag/catcher systems by computational and experimental methods. Experimental testing for intermolecular isopeptide bond formation demonstrated two of the four systems to be functional. For these two systems length and sequence variations of the peptide tags were investigated revealing only a relatively small effect on the efficiency of the reaction. Our study suggests that splitting into tag and catcher moieties is tolerated by a significant portion of the naturally occurring CnaB-domains, thus providing a large reservoir for the design of novel tag/catcher systems.

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Section: Resultsmentioning

confidence: 99%

Probing the potential of CnaB-type domains for the design of tag/catcher systems

et al. 2017

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“…Various techniques are available to decorate VLPs with the antigen(s) of interest [extensively reviewed by Brune and Howarth (14)], such as genetic fusion, chemical derivatization and conjugation, or plug-and-display decoration. This latter technique is based on the isopeptide bond that spontaneously forms between a peptide and its protein couple, derived from specific domains of certain bacterial proteins (15)(16)(17). So far, two binding couples have been developed for vaccine delivery platforms: SpyTag peptide/SpyCatcher protein, derived by splitting the CnaB2 domain of the fibronectin-binding protein FbaB from Streptococcus pyogenes (18); SnoopTag peptide/SnoopCatcher protein, derived by splitting the D4 domain of RrgA adhesin from Streptococcus pneumoniae (19).…”

Section: Introductionmentioning

confidence: 99%

A Universal Plug-and-Display Vaccine Carrier Based on HBsAg VLP to Maximize Effective Antibody Response

Marini

Zhou

et al. 2019

Front. Immunol.

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Development of effective malaria vaccines requires delivery platforms to enhance the immunogenicity and efficacy of the target antigens. This is particularly challenging for transmission-blocking malaria vaccines (TBVs), and specifically for those based on the Pfs25 antigen, that need to elicit very high antibody titers to stop the parasite development in the mosquito host and its transmission. Presenting antigens to the immune system on virus-like particles (VLPs) is an efficient way to improve the quantity and quality of the immune response generated. Here we introduce for the first time a new VLP vaccine platform, based on the well-established hepatitis B surface antigen (HBsAg) fused to the SpyCatcher protein, so that the antigen of interest, linked to the SpyTag peptide, can be easily displayed on it (Plug-and-Display technology). As little as 10% of the SpyCatcher::HBsAg VLPs decorated with Pfs25::SpyTag (molar ratio) induces a higher antibody response and transmission-reducing activity in mice compared to the soluble protein, with 50 and 90% of the VLP coupled to the antigen further enhancing the response. Importantly, using this carrier that is a vaccine antigen itself could be beneficial, as we show that anti-HBsAg IgG antibodies are induced without interfering with the Pfs25-specific immune response generated. Furthermore, pre-existing anti-HBsAg immunity does not affect the antigen-specific response to Pfs25::SpyTag-SpyCatcher::HBsAg, suggesting that these VLPs can have a broad use as a vaccine platform.

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“…Although this bioconjugation method is in its infancy, it has already been deployed by a range of laboratories as a solution to the problems associated with protein ligation. For this system to see further expansion into in vivo applications, continuing optimization of SpyTag and SpyCatcher will need to be explored; indeed, work has already started in this area with the development of traceless tags by using SpyCatcher variants and truncations to reduce immunogenicity as well as orthogonal tags such as SnoopTag and SdyTag . With these improvements, the SpyTag/SpyCatcher system is poised to enable new possibilities in protein synthetic biology.…”

Section: Resultsmentioning

confidence: 99%

Post‐translational Assembly of Protein Parts into Complex Devices by Using SpyTag/SpyCatcher Protein Ligase

2018

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Exploiting the innate modularity of proteins has allowed advances across the fields of synthetic biology and biotechnology. By using standardized protein components as building blocks, complex, multiprotein assemblies with sophisticated functions can be generated; feats previously not possible with strictly genetic‐engineering approaches. The development of strategies for protein assembly is accelerating, pushing the boundaries of protein architecture. SpyTag and SpyCatcher protein ligase is a recent advance in this field that allows plug‐and‐play modularity by harnessing post‐translational protein assembly. Herein, we review the latest applications of this powerful tool including novel enzyme assemblies, modularizing protein display, and the generation of antibody and antibody‐like “devices” by using SpyTag/SpyCatcher technology.

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Kinetic Controlled Tag-Catcher Interactions for Directed Covalent Protein Assembly

Cited by 54 publications

References 28 publications

Probing the potential of CnaB-type domains for the design of tag/catcher systems

Probing the potential of CnaB-type domains for the design of tag/catcher systems

A Universal Plug-and-Display Vaccine Carrier Based on HBsAg VLP to Maximize Effective Antibody Response

Post‐translational Assembly of Protein Parts into Complex Devices by Using SpyTag/SpyCatcher Protein Ligase

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