Intense Pulsed Electric Fields Denature Urease Protein

Urabe, Gen; Katagiri, Toshiaki; Katsuki, S.

doi:10.1089/bioe.2019.0021

Cited by 9 publications

(8 citation statements)

References 33 publications

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“…Summarising, this study demonstrates that EFs of different biologically relevant strengths change S of SARS-CoV-2 both at nanometre and sub-nanometre scales. Considerable changes in the secondary structure of the RBD in the wild type and currently dominant mutants of S occur at field strength orders of magnitude smaller than for most proteins [24][25][26][27][28][29][30][31][32] . We conclude that the spike protein of SARS-CoV-2 (and especially its RBD) is unusually vulnerable to external electric fields.…”

Section: Discussionmentioning

confidence: 97%

“…It has been theoretically predicted and experimentally demonstrated that static and timedependent electric fields (EFs) are capable of inducing conformational changes [25][26][27][28][29][30] or even irreversible damage in proteins [31][32][33] . The fact that extremely intense EFs of strengths larger than 1Volt per nanometre (10 9 V m⁻¹) can denature entire proteins and even break chemical bonds is trivial and of little biological relevance.…”

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confidence: 99%

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The SARS-CoV-2 spike protein is vulnerable to moderate electric fields

et al. 2021

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Most of the ongoing projects aimed at the development of specific therapies and vaccines against COVID-19 use the SARS-CoV-2 spike (S) protein as the main target. The binding of the spike protein with the ACE2 receptor (ACE2) of the host cell constitutes the first and key step for virus entry. During this process, the receptor binding domain (RBD) of the S protein plays an essential role, since it contains the receptor binding motif (RBM), responsible for the docking to the receptor. So far, mostly biochemical methods are being tested in order to prevent binding of the virus to ACE2. Here we show, with the help of atomistic simulations, that external electric fields of easily achievable and moderate strengths can dramatically destabilise the S protein, inducing long-lasting structural damage. One striking field-induced conformational change occurs at the level of the recognition loop L3 of the RBD where two parallel beta sheets, believed to be responsible for a high affinity to ACE2, undergo a change into an unstructured coil, which exhibits almost no binding possibilities to the ACE2 receptor. We also show that these severe structural changes upon electric-field application also occur in the mutant RBDs corresponding to the variants of concern (VOC) B.1.1.7 (UK), B.1.351 (South Africa) and P.1 (Brazil). Remarkably, while the structural flexibility of S allows the virus to improve its probability of entering the cell, it is also the origin of the surprising vulnerability of S upon application of electric fields of strengths at least two orders of magnitude smaller than those required for damaging most proteins. Our findings suggest the existence of a clean physical method to weaken the SARS-CoV-2 virus without further biochemical processing. Moreover, the effect could be used for infection prevention purposes and also to develop technologies for in-vitro structural manipulation of S. Since the method is largely unspecific, it can be suitable for application to other mutations in S, to other proteins of SARS-CoV-2 and in general to membrane proteins of other virus types.

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Section: Discussionmentioning

confidence: 97%

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confidence: 99%

The SARS-CoV-2 spike protein is vulnerable to moderate electric fields

et al. 2021

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“…Applying 10 6 V/cm on a protein crystal results in faint structural changes . However, experimentally, lower electric fields can have deleterious consequences on protein structure or activity, either fields of 5–50 × 10 3 V/cm applied during a few microseconds − ,, or 50–500 V/cm applied through minutes to hours. , Frictional forces due to electrophoretic motion of the corresponding purified proteins might be the cause of these observations . These scenarios do not correspond to the common pulsing protocols used for electroporation delivery.…”

Section: Methodsmentioning

confidence: 99%

In-Cell Structural Biology by NMR: The Benefits of the Atomic Scale

Theillet

2022

Chem. Rev.

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In-cell structural biology aims at extracting structural information about proteins or nucleic acids in their native, cellular environment. This emerging field holds great promises and is already providing new facts and outlooks of interest at both fundamental and applied levels. NMR spectroscopy has important contributions on this stage: it brings information on a broad variety of nuclei at the atomic scale, which ensures its great versatility and uniqueness. Here, we detail the methods, the fundamental knowledge and the applications in biomedical engineering related to in-cell NMR. We finally propose a brief overview of the main other techniques in the field (EPR, smFRET, cryo-ET…) to draw some advisable developments for in-cell NMR. In the era of large-scale screenings and deep learning, both accurate and qualitative experimental evidences are as essential as ever to understand the interior life of cells. In-cell structural biology by NMR spectroscopy can generate such a knowledge, and it does so at the atomic scale. This review is meant to deliver comprehensive but accessible information, with advanced technical details and reflections on the methods, the nature of the results and the future of the field. CONTENTS 5.1.1. In-cell EPR 5.1.2. In-cell FRET microscopy 5.1.3. Mass-spectrometry 5.1.4. Cryo-ET 5.2. The specific benefits of in-cell NMR 5.3. The future technical challenges of in-cell NMR 6. Conclusion Author Information

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“…Although providing exquisite atomistic details, that experiment was performed on a protein crystal, so its relevance to proteins under physiological condition (in solution) is limited. In solution, Urabe et al have shown that a train (500) of 5 ns, 3 MV/m electric pulses can physically denature urease [55] and a train (1000) of 1 ns, 200 MV/m pulses promotes the disintegration of transthyretin amyloid [56] . In addition, in our recent pioneering studies, we showed that an intense electric field has multiple effects on a single tubulin in vitro and in silico .…”

Section: Introductionmentioning

confidence: 99%