Tuning a coiled-coil hydrogel <i>via</i> computational design of supramolecular fiber assembly

Britton, Dustin; Meleties, Michael; Liu, Chengliang; Jia, Sihan; Mahmoudinobar, Farbod; Renfrew, P. Douglas; Bonneau, Richard; Montclare, Jin Kim

doi:10.1039/d2me00153e

Cited by 5 publications

(45 citation statements)

References 43 publications

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“…The Rosetta Relax protocol was used on protein sequences maintaining the symmetry of COMPcc (PDB: 3V2P) with the all-atom energy score function . Q2 and Q3 were designed previously, where Q2 has been characterized as a hydrogel and fiber , and Q3 as a fiber . PDB2QR and APBS were used to calculate the electrostatic potential of surface residues, EE bcf , for the N- and C-termini.…”

Section: Methodsmentioning

confidence: 99%

“…Binary sol–gel behavior was assessed using tube inversion . Immediately after concentration to 2 mM, 150 μL of Q hydrogel variants were incubated in 2 mL microtubes at 4 °C for 2 weeks.…”

Section: Methodsmentioning

confidence: 99%

“…Among the coiled–coil hydrogel sequences studied, Q5 undergoes a complete sol–gel transition within 24 h, possessing a critical gelation time, t c , of 11.5 h, a greater than 2-fold increase to previously reported coiled–coiled hydrogels . For fiber formation, transmission electron microscopy (TEM) suggests a mechanism predominantly dependent on end-to-end stacking .…”

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: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

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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“…Rationale and Protein Synthesis. Q2 TFL was designed for greater thermostability possessing 9 leucines, compared to 7 in Q TFL , which was confirmed by a higher Rosetta Score 39,40 with the aim of creating a fluorinated fiber capable of curcumin (CCM) encapsulation (Figure 1a). Q2 TFL was generated by residue-specific noncanonical amino acid incorporation of trifluoroleucine (TFL) using leucine auxotrophic LAM1000 E. coli cells.…”

Section: ■ Results and Discussionmentioning

confidence: 99%

“…Q2 TFL was designed for greater thermostability possessing 9 leucines, compared to 7 in Q TFL , which was confirmed by a higher Rosetta Score , with the aim of creating a fluorinated fiber capable of curcumin (CCM) encapsulation (Figure a). Q2 TFL was generated by residue-specific noncanonical amino acid incorporation of trifluoroleucine (TFL) using leucine auxotrophic LAM1000 E. coli cells. , Protein expression (Figure b) and purification (Figure c) were assessed by 12% SDS-PAGE gels showing protein bands at a molecular weight of 6.97 kDa for Q2 TFL .…”

Section: Resultsmentioning

confidence: 99%

Protein-Engineered Fibers For Drug Encapsulation Traceable via ¹⁹F Magnetic Resonance

Britton,

Legocki,

Aristizabal

et al. 2023

ACS Appl. Nano Mater.

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Theranostic materials research is experiencing rapid growth driven by the interest in integrating both therapeutic and diagnostic modalities. These materials offer the unique capability to not only provide treatment but also track the progression of a disease. However, to create an ideal theranostic biomaterial without compromising drug encapsulation, diagnostic imaging must be optimized for improved sensitivity and spatial localization. Herein, we create a protein-engineered fluorinated coiled-coil fiber, Q2 TFL , capable of improved sensitivity to 19 F magnetic resonance spectroscopy (MRS) detection. Leveraging residue-specific noncanonical amino acid incorporation of trifluoroleucine (TFL) into the coiled-coil, Q2, which self-assembles into nanofibers, we generate Q2 TFL . We demonstrate that fluorination results in a greater increase in thermostability and 19 F magnetic resonance detection compared to the nonfluorinated parent, Q2. Q2 TFL also exhibits linear ratiometric 19 F MRS thermoresponsiveness, allowing it to act as a temperature probe. Furthermore, we explore the ability of Q2 TFL to encapsulate the anti-inflammatory small molecule, curcumin (CCM), and its impact on the coiled-coil structure. Q2 TFL also provides hyposignal contrast in 1 H MRI, echogenic signal with high-frequency ultrasound and sensitive detection by 19 F MRS in vivo illustrating fluorination of coiled-coils for supramolecular assembly and their use with 1 H MRI, 19 F MRS and high frequency ultrasound as multimodal theranostic agents.

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Exosome Loaded Protein Hydrogel for Enhanced Gelation Kinetics and Wound Healing

Britton,

Almanzar,

Xiao

et al. 2024

ACS Appl. Bio Mater.

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Exosomes are being increasingly explored in biomedical research for wound healing applications. Exosomes can improve blood circulation and endocrine signaling, resulting in enhanced cell regeneration. However, exosome treatments suffer from low retention and bioavailability of exosomes at the wound site. Hydrogels are a popular tool for drug delivery due to their ability to encapsulate drugs in their network and allow for targeted release. Recently, hydrogels have proven to be an effective method to provide increased rates of wound healing when combined with exosomes that can be applied noninvasively. We have designed a series of single-domain protein-based hydrogels capable of physical cross-linking and upper critical solution temperature (UCST) behavior. Hydrogel variant Q5, previously designed with improved UCST behavior and a significantly enhanced gelation rate, is selected as a candidate for encapsulation release of exosomes dubbed Q5Exo. Q5Exo exhibits low critical gelation times and significant decreases in wound healing times in a diabetic mouse wound model showing promise as an exosome-based drug delivery tool and for future hybrid, noninvasive protein-exosome design.

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Tuning a coiled-coil hydrogel via computational design of supramolecular fiber assembly

Abstract: Previously reported, Q, is a thermoresponsive coiled-coil protein capable of higher-order supramolecular assembly into fibers and hydrogels with upper critical solution temperature (UCST) behavior. Here, we introduce a new coiled-coil...

Cited by 5 publications

References 43 publications

Computational Prediction of Coiled–Coil Protein Gelation Dynamics and Structure

Computational Prediction of Coiled–Coil Protein Gelation Dynamics and Structure

Protein-Engineered Fibers For Drug Encapsulation Traceable via ¹⁹F Magnetic Resonance

Exosome Loaded Protein Hydrogel for Enhanced Gelation Kinetics and Wound Healing

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Tuning a coiled-coil hydrogel via computational design of supramolecular fiber assembly

Abstract: Previously reported, Q, is a thermoresponsive coiled-coil protein capable of higher-order supramolecular assembly into fibers and hydrogels with upper critical solution temperature (UCST) behavior. Here, we introduce a new coiled-coil...

Cited by 5 publications

References 43 publications

Computational Prediction of Coiled–Coil Protein Gelation Dynamics and Structure

Computational Prediction of Coiled–Coil Protein Gelation Dynamics and Structure

Protein-Engineered Fibers For Drug Encapsulation Traceable via 19F Magnetic Resonance

Exosome Loaded Protein Hydrogel for Enhanced Gelation Kinetics and Wound Healing

Contact Info

Product

Resources

About

Protein-Engineered Fibers For Drug Encapsulation Traceable via ¹⁹F Magnetic Resonance