Jin Kim Montclare

Protein folding dynamics emerge from subtle changes in intramolecular interactions, where hydration shapes free energy landscapes and influences conformational ensembles and supramolecular assembly. While these shape-shifting principles have been primarily explored in well-behaved systems with sharp transitions, their applicability to multidomain protein assemblies, particularly those containing partially folded or disordered regions, remains unclear. Here, we investigate how a single leucine-to-alanine mutation modulates the hierarchical interplay between folding, micelle formation, and macroscopic biomaterial behavior using two diblock protein block copolymers (BCPs): CE and C_L44AE. In solution, both BCPs exhibit spectral features consistent with their C/C_L44A monoblocks, with CE showing α-helical features and C_L44AE more disordered ones. Upon dehydration, however, both systems display increased structural order, with CE favoring β-sheets and C_L44AE adopting α-helical content. At the mesoscale, both BCPs assemble into amphiphilic micelles, but CE forms smaller, densely packed micelles, whereas C_L44AE generates larger micelles that template film networks of varying pore size depending on their packing density. These differences lead to distinct solvation behavior, with C_L44AE exhibiting greater uptake of predominantly mobile water, approximately twice that of CE, culminating in an actuation energy density of 2,043 kJ/m³, the highest reported for protein BCPs to date. Despite this increased uptake, C_L44AE shows reduced mechanical relaxation and a rougher DSC/TGA thermogram, indicative of a broader conformational ensemble compared to CE. Comparisons to monoblocks (C, C_L44A, E) and previously reported triblocks (CEC, C_L44AEC_L44A) further highlight the importance of asymmetric diblock architectures in promoting hydration-sensitive assemblies, laying the rational design groundwork for future high-performance, water-responsive protein biomaterials.

Protein‐based nanocarriers bear highly desirable properties such as biodegradability and the ability to facilitate passage through biological barriers such as the blood–brain‐barrier. Using modular protein engineering, we develop a strategy for iteratively improving the delivery efficacy of hydrophobic small molecules for the treatment of glioblastoma multiforme (GBM). By increasing the multiplicity m of the coiled‐coil and RGD peptide targeting regions from 1 to 2, we can increase both the hydrodynamic micellar size and drug loading capacity of the targeted multidomain protein assembly (TMPA) relative to its predecessor thermoresponsive assembled protein (TRAP). An upper limit of m is likely determined by steric interactions. TMPA shows a 1.7‐fold increase in doxorubicin (Dox) encapsulation compared to TRAP and demonstrates a 1.3‐fold improvement in uptake by U87 human GBM cells. Near‐infrared (NIR) dye‐labelled TMPA (NIR‐TMPA) is intravenously administered to mice orthotopically implanted with GBM cells and to control mice. Pharmacokinetic analysis using a 2‐compartment pharmacokinetic model reveals a significantly prolonged distribution‐phase (short‐phase) half‐life in tumor‐bearing mice compared to control, while the elimination‐phase (slow‐phase) half‐life remains comparable between groups. This suggests altered early‐phase kinetics likely due to tumor‐associated sequestration or retention. The resulting increased area under the concentration‐time curve (AUC) in tumor‐bearing mice supports enhanced accumulation or slower clearance. Ex vivo fluorescence imaging of organs and 3D reconstructions of whole mouse heads further corroboratesa preferential localization of NIR‐TMPA in tumor regions. These findings highlight the potential of TMPA and its future derivatives for targeted GBM therapy.

Protein‐based nanocarriers bear highly desirable properties such as biodegradability and the ability to facilitate passage through biological barriers such as the blood–brain‐barrier. Using modular protein engineering, we develop a strategy for iteratively improving the delivery efficacy of hydrophobic small molecules for the treatment of glioblastoma multiforme (GBM). By increasing the multiplicity m of the coiled‐coil and RGD peptide targeting regions from 1 to 2, we can increase both the hydrodynamic micellar size and drug loading capacity of the targeted multidomain protein assembly (TMPA) relative to its predecessor thermoresponsive assembled protein (TRAP). An upper limit of m is likely determined by steric interactions. TMPA shows a 1.7‐fold increase in doxorubicin (Dox) encapsulation compared to TRAP and demonstrates a 1.3‐fold improvement in uptake by U87 human GBM cells. Near‐infrared (NIR) dye‐labelled TMPA (NIR‐TMPA) is intravenously administered to mice orthotopically implanted with GBM cells and to control mice. Pharmacokinetic analysis using a 2‐compartment pharmacokinetic model reveals a significantly prolonged distribution‐phase (short‐phase) half‐life in tumor‐bearing mice compared to control, while the elimination‐phase (slow‐phase) half‐life remains comparable between groups. This suggests altered early‐phase kinetics likely due to tumor‐associated sequestration or retention. The resulting increased area under the concentration‐time curve (AUC) in tumor‐bearing mice supports enhanced accumulation or slower clearance. Ex vivo fluorescence imaging of organs and 3D reconstructions of whole mouse heads further corroboratesa preferential localization of NIR‐TMPA in tumor regions. These findings highlight the potential of TMPA and its future derivatives for targeted GBM therapy.

Photoconductive nanomaterials are critical in the development of advanced optoelectronics, which span photovoltaics to biomedicine. These materials are often derived from inorganic, molecular, and polymeric crystalline semiconductors that facilitate charge transfer upon absorption of ultraviolet and near infrared light. These types of materials are broadened to include single‐domain protein‐based nanofibers. Coiled‐coil protein domains found in nature transport electrons when possessing pi‐stacked aromatic rings and undergo supramolecular assembly into fibers. Whereas conductive protein nanofibers have previously been leveraged for high‐resistance organic materials, a conductive helical assembled fiber (CHAF) is reported, composed of phenylalanines in the core capable of photoresponsivity. It is demonstrated that CHAF and a leucine‐replaced CHAF control (L‐CHAF) exhibit distinct photoconductive responses to near infrared (NIR) light. The sequence‐controlled system reported here illustrates photoconductive protein fibers as a promising platform for the next generation of new biocompatible nanomaterials.

Gamified simulations, integrating gameplay into education, cater to younger learners’ digital preferences and align with Next Generation Science Standards. Current virtual modules focus on advanced high school classes, leaving a gap for middle school students. This study investigated the impact of substituting recitations in a 6th-grade ecology class with Feed the Dingo , a gamified module. Through quizzes evaluating academic performance and free-response surveys to gauge students’ attitudes, the module appeared to enhance intuitive understanding of core ecological concepts (e.g., ecosystems, food webs, biodiversity, etc.), resulting in commendable academic achievement and positive feedback. Such simulations serve as valuable supplements for K-12 lesson planning.

Rheological modifiers are an important component in the development of skin cream (SC) chassis for personal skin care products (PSCPs). The viscous behavior of a PSCP is critical to its effectiveness where its uniformity and material strength impact its processing, storage, and delivery of active ingredients. Due to the mildly acidic environment of the skin, PSCPs require a SC that will assist in maintaining their material strength at low pHs. We have investigated a coiled-coil protein hydrogel system for the ability to possess pH-responsiveness, where physical cross-linking and material strength is controlled by pH relative to the isoelectric point (pI) of the protein. We recently designed a coiled-coil protein hydrogel variant, Q5, which possesses a relatively low pI that we hypothesized to have improved supramolecular assembly into a hydrogel at acidic conditions. We demonstrate that Q5 can retain a partial solution-to-gel transition at pH 6.0 and acts as a soft hydrogel by rheology. We further tested Q5 to act as a rheological modifier in a standard SC at pH 6.0 and pH 8.0 to test conditions mediated by pH changes in the skin environment. Q5 reveals the ability to uniquely increase material strength at low pH in comparison to a standard rheological modifier like hydroxyethyl cellulose (HEC), suggesting modular protein-based coiled-coil rheological modifiers can be used in PSCPs.