Signal amplification by a self-assembled biosensor system designed on the principle of dockerin–cohesin interactions in a cellulosome complex

Hyeon, Jeong Eun; Kang, Dae Hee; Han, Sung Ok

doi:10.1039/c4an00856a

“…[14,24] We provide here a brief overview of the current state of the art in molecular-level knowledge of how these structures function, to provide context and to illustrate the broad lessons available to materials science from (evolved) biological nanostructures. [25] See, for example, the self-assembled biosensor described by Han et al [26] . These molecular machines operate at scales between 0.1 and 100 nanometers, exploiting nanoscale physics, in particular Brownian motion and van der Waals sticking forces, to manipulate individual atoms in order to process (and manufacture) materials from the bottom up.…”

Section: Biocatalystsmentioning

confidence: 99%

Nanoscale Engineering of Designer Cellulosomes

Gunnoo

¹

,

Cazade

²

,

Galera‐Prat

³

et al. 2016

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Biocatalysts showcase the upper limit obtainable for high-speed molecular processing and transformation. Efforts to engineer functionality in synthetic nanostructured materials are guided by the increasing knowledge of evolving architectures, which enable controlled molecular motion and precise molecular recognition. The cellulosome is a biological nanomachine, which, as a fundamental component of the plant-digestion machinery from bacterial cells, has a key potential role in the successful development of environmentally-friendly processes to produce biofuels and fine chemicals from the breakdown of biomass waste. Here, the progress toward so-called "designer cellulosomes", which provide an elegant alternative to enzyme cocktails for lignocellulose breakdown, is reviewed. Particular attention is paid to rational design via computational modeling coupled with nanoscale characterization and engineering tools. Remaining challenges and potential routes to industrial application are put forward.

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“…, 2012 ) and existence of different specificity groups ( Haimovitz et al ., 2008 ) make them very suitable to generate well-defined multi-protein complexes generally known as designer cellulosomes ( Bayer et al ., 1994 ; Fierobe et al ., 2001 ). This technology has been shown to be an invaluable tool to study cellulosome activity, and the principle has also been applied to other fields such as biosensor development ( Hyeon et al ., 2014 ), as well as to improve of reactions taking advantage of the proximity induced by assembly on the scaffold ( You et al ., 2012 ; Liu et al ., 2013 ; You and Zhang, 2014 ).…”

Section: Introductionmentioning

confidence: 99%

Protein engineering approach to enhance activity assays of mono-ADP-ribosyltransferases through proximity

Galera‐Prat

¹

,

Alaviuhkola

²

,

Alanen

³

et al. 2022

Protein Engineering, Design and Selection

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Human mono-ADP-ribosylating PARP enzymes have been linked to several clinically relevant processes and many of these PARPs have been suggested as potential drug targets. Despite recent advances in the field, efforts to discover such compounds have been hindered by the lack of tools to rapidly screen for high potency compounds and profile them against the different PARP enzymes of the ARTD family. We here expanded the methods and engineered mono-ART catalytic fragments to be incorporated into a cellulosome-based octavalent scaffold. Compared to the free enzymes, the scaffold-based system results in an improved activity for the tested PARPs due to improved solubility, stability and the proximity of the catalytic domains, altogether boosting their activity beyond 10-fold in the case of PARP12. This allows us to measure their enhanced activity using a simple and easily accessible homogeneous NAD+ conversion assay, facilitating its automation to reduce the assay volume and lowering the assay costs. The approach will enable the discovery of more potent compounds due to increased assay sensitivity and it can be applied to compound screening campaigns as well as inhibitor profiling.

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“…The modularity of these components, together with their high affinity (Slutzki et al, 2012) and existence of different specificity groups (Haimovitz et al, 2008) make them very suitable to generate well-defined multi-protein complexes generally known as designer cellulosomes (Bayer et al, 1994; Fierobe et al, 2001). This technology has been shown to be an invaluable tool to study cellulosome activity and the principle has also been applied to other fields such as biosensor development (Hyeon et al, 2014), as well as to improve of reactions taking advantage of the proximity induced by assembly on the scaffold (You et al, 2012; Liu et al, 2013; You and Zhang, 2014).…”

Section: Introductionmentioning

confidence: 99%

Protein engineering approach to enhance activity assays of mono-ADP-ribosyltransferases through proximity

Galera‐Prat

¹

,

Alaviuhkola

²

,

Alanen

³

et al. 2022

Preprint

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Human mono-ADP-ribosylating PARP enzymes have been linked to several clinically relevant processes and many of these PARPs have been suggested as potential drug targets. Despite recent advances in the field, efforts to discover such compounds have been hindered by the lack of tools to rapidly screen for high potency compounds and profile them against the different PARP enzymes of the ARTD family. We here expanded the methods and engineered mono-ART catalytic fragments to be incorporated into a cellulosome-based octavalent scaffold. Compared to the free enzymes, the scaffold-based system results in an improved activity for the tested PARPs due to improved solubility, stability and the proximity of the catalytic domains, altogether boosting their activity beyond 10-fold in the case of PARP12. This allows us to measure their enhanced activity using a simple and easily accessible homogeneous NAD+ conversion assay, facilitating its automation to reduce the assay volume and lowering the assay costs. The approach will enable the discovery of more potent compounds due to increased assay sensitivity and it can be applied to compound screening campaigns as well as inhibitor profiling.

show abstract

Signal amplification by a self-assembled biosensor system designed on the principle of dockerin–cohesin interactions in a cellulosome complex

Cited by 15 publications

References 16 publications

Nanoscale Engineering of Designer Cellulosomes

Nanoscale Engineering of Designer Cellulosomes

Protein engineering approach to enhance activity assays of mono-ADP-ribosyltransferases through proximity

Protein engineering approach to enhance activity assays of mono-ADP-ribosyltransferases through proximity

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