Fluorescent azobenzene-confined coiled-coil mesofibers

Punia, Kamia; Britton, Dustin; Hüll, Katharina; Liang, Yin; Wang, Yifei; Renfrew, P. Douglas; Gilchrist, M. Lane; Bonneau, Richard; Trauner, Dirk; Montclare, Jin Kim

doi:10.1039/d2sm01578a

Cited by 3 publications

(3 citation statements)

References 32 publications

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“…Fluorescent recovery after photobleaching (FRAP) is measured by photobleaching using a 405 nm laser and measuring the spot intensity over time. We have previously confirmed this behavior in our Q protein in a recent characterization . Using the correlation between autofluorescence and fibrilization, we measured the relative autofluorescence of our coiled–coil protein hydrogels.…”

Section: Resultsmentioning

confidence: 99%

“…We have previously confirmed this behavior in our Q protein in a recent characterization. 18 Using the correlation between autofluorescence and fibrilization, we measured the relative autofluorescence of our coiled–coil protein hydrogels. Interestingly, the fluorescence intensities of the hydrogels diminished nearest the center of the laser, indicating a loss of fibrilization over time.…”

Section: Resultsmentioning

confidence: 99%

“…Previously, we have designed upper critical solution temperature (UCST) coiled–coil protein hydrogels by targeting electrostatic interactions between the N-terminus possessing a positively charged surface patch and the C-terminus possessing a negatively charged surface patch. , Notably, significant differences in physically cross-linked fiber diameters and their respective critical gelation times were reported for proteins designed to possess a lower electrostatic potential and improved thermostability . While protein domain folding − or MD software has been used to assess stability of coiled–coil self-assembly, a broader gap exists in sequence-structure-gelation relationships. Recently, we have established a quantitative relationship between fiber diameters and the electrostatic potential difference between the N- and C- termini of surface residues in the b, c, and f helical wheel positions (ΔEE bcf ) as a metric to understand the role of electrostatic interactions on supramolecular nanofiber assembly …”

Section: Introductionmentioning

confidence: 99%

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Computational Prediction of Coiled–Coil Protein Gelation Dynamics and Structure

Britton,

Christians,

Liu

et al. 2023

Biomacromolecules

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Protein hydrogels represent an important and growing biomaterial for a multitude of applications, including diagnostics and drug delivery. We have previously explored the ability to engineer the thermoresponsive supramolecular assembly of coiled−coil proteins into hydrogels with varying gelation properties, where we have defined important parameters in the coiled−coil hydrogel design. Using Rosetta energy scores and Poisson−Boltzmann electrostatic energies, we iterate a computational design strategy to predict the gelation of coiled−coil proteins while simultaneously exploring five new coiled−coil protein hydrogel sequences. Provided this library, we explore the impact of in silico energies on structure and gelation kinetics, where we also reveal a range of blue autofluorescence that enables hydrogel disassembly and recovery. As a result of this library, we identify the new coiled−coil hydrogel sequence, Q5, capable of gelation within 24 h at 4 °C, a more than 2-fold increase over that of our previous iteration Q2. The fast gelation time of Q5 enables the assessment of structural transition in real time using small-angle X-ray scattering (SAXS) that is correlated to coarse-grained and atomistic molecular dynamics simulations revealing the supramolecular assembling behavior of coiled−coils toward nanofiber assembly and gelation. This work represents the first system of hydrogels with predictable self-assembly, autofluorescent capability, and a molecular model of coiled−coil fiber formation.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Computational Prediction of Coiled–Coil Protein Gelation Dynamics and Structure

Britton,

Christians,

Liu

et al. 2023

Biomacromolecules

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Dual coiled-coil protein domain mimic and drug delivery vehicle for SARS-CoV-2

Britton,

Liu,

Jia

et al. 2024

Biochemical Engineering Journal

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Design of Coiled-Coil Protein Nanostructures for Therapeutics and Drug Delivery

Britton,

Sun,

Renfrew

et al. 2024

Annual Review of Chemical and Biomolecular Engineering

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Coiled-coil protein motifs have become widely employed in the design of biomaterials. Some of these designs have been studied for use in drug delivery due to the unique ability of coiled-coils to impart stability, oligomerization, and supramolecular assembly. To leverage these properties and improve drug delivery, release, and targeting, a variety of nano- to mesoscale architectures have been adopted. Coiled-coil drug delivery and therapeutics have been developed by using the coiled-coil alone, designing for higher-order assemblies such as fibers and hydrogels, and combining coiled-coil proteins with other biocompatible structures such as lipids and polymers. We review the recent development of these structures and the design criteria used to generate functional proteins of varying sizes and morphologies.

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Fluorescent azobenzene-confined coiled-coil mesofibers

Abstract: Fluorescent protein biomaterials have important applications such as bioimaging in pharmacological studies. Self-assembly of proteins, especially into fibrils, is known to produce fluorescence in the blue band. Capable of self-assembly...

Cited by 3 publications

References 32 publications

Computational Prediction of Coiled–Coil Protein Gelation Dynamics and Structure

Computational Prediction of Coiled–Coil Protein Gelation Dynamics and Structure

Dual coiled-coil protein domain mimic and drug delivery vehicle for SARS-CoV-2

Design of Coiled-Coil Protein Nanostructures for Therapeutics and Drug Delivery

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