Engineering peroxiredoxin 3 to facilitate control over self-assembly

Conroy, Frankie; Rossi, Tatiana Spatola; Ashmead, Helen; Crowther, Jennifer M.; Mitra, Alok K.; Gerrard, Juliet A.

doi:10.1016/j.bbrc.2019.03.032

Cited by 3 publications

(6 citation statements)

References 29 publications

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“…These results account for the high affinity of Ni 2+ ions for the polyHis sequences ( K d = 15 nM) located at the ring pore that become driving forces to achieve stacking into tubes, acting as polymerizing agents increasing the local concentration of the rings. In fact, both the top and bottom surfaces of the polyHis-tagged rings are suitable environments to bind divalent metal ions within a binding surface of ∼130 nm 2 . , Similarly, the N-terminal polyHis-tagged hPrxIII constitutively forms short stacks at pH 7.2, even in the presence of EDTA . Interestingly, changing the length of the tag affects the tube elongation with the number of rings incorporated within the stacks being 2–10, 2–66, and 4–100 for tags containing 2, 4, and 6 histidine residues, respectively .…”

Section: Peroxiredoxin-based Bionanotechnologymentioning

confidence: 99%

“…In fact, both the top and bottom surfaces of the polyHis-tagged rings are suitable environments to bind divalent metal ions within a binding surface of ∼130 nm 2 . , Similarly, the N-terminal polyHis-tagged hPrxIII constitutively forms short stacks at pH 7.2, even in the presence of EDTA . Interestingly, changing the length of the tag affects the tube elongation with the number of rings incorporated within the stacks being 2–10, 2–66, and 4–100 for tags containing 2, 4, and 6 histidine residues, respectively . This effect can be ascribable to two factors modulated by the elongation of the N-termini: (1) π–π stacking of the histidines’ imidazole groups between adjoining rings and/or (2) coordination binding of monovalent and/or divalent metal ions, not efficiently sequestered by EDTA, to the polyHis tags resulting in bridging bonds between rings.…”

Section: Peroxiredoxin-based Bionanotechnologymentioning

confidence: 99%

“…Interestingly, in most cases changing these factors facilitates tuning the morpheein behavior and the interaction with nanomaterials. Evidence is presented for how to harness the oligomeric transitions of Prxs in solution from rings ,,,,,− to hollow colloidal and surface-tethered tubes ,,,,, as well as cage-like particles and catenanes. , Studies also show methods to improve such complexes making them suitable tectons for building nanostructures such as 2D protein arrays and nanoribbon-based 3D hydrogels . Their derivatives can be coupled to optoelectronic compounds for building transistor devices, 0D and 1D arrays of colloidal metal and mineralized NPs with electrical polarizability , and 1D SERS-active metal NPs clusters .…”

Section: Conclusion and Future Perspectivesmentioning

confidence: 99%

“…78,79 A different approach relies on ad hoc amino acid mutations that prevent stacking of the rings into HMW species by destabilizing the ring−ring R-interface without hampering the ring assembly. For instance, the N-terminal polyHis-tagged mutants His164Glu, His164Ala, and Thr163Val of hPrxIII form rings at pH 7.2 in the presence of the metal chelator ethylenediamine tetraacetic acid (EDTA); conversely, the tagged wild-type protein can still stack, 80 in contrast to other results (see above). 29 To explain such unusual behavior, the authors considered the involvement of π−π stacking between the imidazole groups of the polyHis tags from adjoining hPrxIII rings and the possibility that the protein, as an ATPindependent holdase activity, may recognize the 33-aminoacid-long polyHis tag as unfolded peptide thus triggering HMW tube self-assembly, aided by electrostatic interactions between the positively charged tags and the negatively charged α2-helices at the R-interface.…”

Section: Peroxiredoxin-based Bionanotechnologymentioning

confidence: 99%

“…46,84 Similarly, the N-terminal polyHis-tagged hPrxIII constitutively forms short stacks at pH 7.2, even in the presence of EDTA. 80 Interestingly, changing the length of the tag affects the tube elongation with the number of rings incorporated within the stacks being 2−10, 2− 66, and 4−100 for tags containing 2, 4, and 6 histidine residues, respectively. 80 This effect can be ascribable to two factors modulated by the elongation of the N-termini: (1) π−π stacking of the histidines' imidazole groups between adjoining rings and/or (2) coordination binding of monovalent and/or divalent metal ions, not efficiently sequestered by EDTA, to the polyHis tags resulting in bridging bonds between rings.…”

Section: Peroxiredoxin-based Bionanotechnologymentioning

confidence: 99%

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Taking Advantage of the Morpheein Behavior of Peroxiredoxin in Bionanotechnology

et al. 2021

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Morpheeins are proteins that reversibly assemble into different oligomers, whose architectures are governed by conformational changes of the subunits. This property could be utilized in bionanotechnology where the building of nanometric and new high-ordered structures is required. By capitalizing on the adaptability of morpheeins to create patterned structures and exploiting their inborn affinity toward inorganic and living matter, “bottom-up” creation of nanostructures could be achieved using a single protein building block, which may be useful as such or as scaffolds for more complex materials. Peroxiredoxins represent the paradigm of a morpheein that can be applied to bionanotechnology. This review describes the structural and functional transitions that peroxiredoxins undergo to form high-order oligomers, e.g., rings, tubes, particles, and catenanes, and reports on the chemical and genetic engineering approaches to employ them in the generation of responsive nanostructures and nanodevices. The usefulness of the morpheeins’ behavior is emphasized, supporting their use in future applications.

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Section: Peroxiredoxin-based Bionanotechnologymentioning

confidence: 99%

Section: Peroxiredoxin-based Bionanotechnologymentioning

confidence: 99%

Section: Conclusion and Future Perspectivesmentioning

confidence: 99%

Section: Peroxiredoxin-based Bionanotechnologymentioning

confidence: 99%

Section: Peroxiredoxin-based Bionanotechnologymentioning

confidence: 99%

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Taking Advantage of the Morpheein Behavior of Peroxiredoxin in Bionanotechnology

et al. 2021

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Peroxiredoxin Proteins as Building Blocks for Nanotechnology

Conroy

Yewdall²

2019

Methods in Molecular Biology

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Fluorescence Lifetime Phasor Analysis of the Decamer–Dimer Equilibrium of Human Peroxiredoxin 1

Ferrer-Sueta

Denicola

2022

IJMS

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Protein self-assembly is a common feature in biology and is often required for a myriad of fundamental processes, such as enzyme activity, signal transduction, and transport of solutes across membranes, among others. There are several techniques to find and assess homo-oligomer formation in proteins. Naturally, all these methods have their limitations, meaning that at least two or more different approaches are needed to characterize a case study. Herein, we present a new method to study protein associations using intrinsic fluorescence lifetime with phasors. In this case, the method is applied to determine the equilibrium dissociation constant (KD) of human peroxiredoxin 1 (hPrx1), an efficient cysteine-dependent peroxidase, that has a quaternary structure comprised of five head-to-tail homodimers non-covalently arranged in a decamer. The hPrx1 oligomeric state not only affects its activity but also its association with other proteins. The excited state lifetime of hPrx1 has distinct values at high and low concentrations, suggesting the presence of two different species. Phasor analysis of hPrx1 emission lifetime allowed for the identification and quantification of hPrx1 decamers, dimers, and their mixture at diverse protein concentrations. Using phasor algebra, we calculated the fraction of hPrx1 decamers at different concentrations and obtained KD (1.1 × 10−24 M4) and C0.5 (1.36 μM) values for the decamer–dimer equilibrium. The results were validated and compared with size exclusion chromatography. In addition, spectral phasors provided similar results despite the small differences in emission spectra as a function of hPrx1 concentration. The phasor approach was shown to be a highly sensitive and quantitative method to assess protein oligomerization and an attractive addition to the biophysicist’s toolkit.

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Engineering peroxiredoxin 3 to facilitate control over self-assembly

Cited by 3 publications

References 29 publications

Taking Advantage of the Morpheein Behavior of Peroxiredoxin in Bionanotechnology

Taking Advantage of the Morpheein Behavior of Peroxiredoxin in Bionanotechnology

Peroxiredoxin Proteins as Building Blocks for Nanotechnology

Fluorescence Lifetime Phasor Analysis of the Decamer–Dimer Equilibrium of Human Peroxiredoxin 1

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