1. ABOUT THE DATASET -------------------- Title: Capturing the Impact of Protein Unfolding on the Dynamic Assembly of Protein Networks Creator(s): Matt D G Hughes[1], Sophie Cussons [2,3], Ahmad Boroumand [1], Arwen I I Tyler [4], David J Brockwell [2,3], and Lorna Dougan [1,2] Organisation(s): [1] School of Physics and Astronomy, Faculty of Engineering and Physical Sciences, University of Leeds, UK [2] Astbury Centre for Structural Molecular Biology, University of Leeds, UK [3] School of Molecular and Cellular Biology, Faculty of Biological Sciences, University of Leeds, UK [4] School of Food Science and Nutrition, Faculty of Environment, University of Leeds, UK Rights-holder(s):Unless otherwise stated, Copyright 2025 University of Leeds Publication Year: 2025 Description: The rapid assembly of molecular or nanoscale building blocks into extended arrays is crucial to the construction of functional networks in vivo and in vitro, depending on various factors. One factor seldom considered is the dynamic changes of the building block shape. Folded protein building blocks offer a unique system to investigate dynamic shape changes due to their intrinsic ability to change from a compact and specific folded structure to an extended unfolded structure in response to a perturbation such as force. Here, we use photochemically cross-linked folded protein hydrogels constructed from force labile protein building blocks as a model dynamic shape-changing network system and characterise them by combining time-resolved rheology and small-angle x-ray scattering (SAXS). This approach probes both the load-bearing network structures, using rheology, and network architectures, using SAXS, thereby providing a cross-length scale understanding of the network formation. We propose a triple assembly model for the structural evolution of networks constructed from force labile protein building block consisting of: primary formation where monomeric folded proteins create the preliminary protein network scaffold; a subsequent secondary formation phase, where larger oligomers of protein diffuse to join the preliminary network scaffold; and finally in situ unfolding and relaxation which leads to the mature network structure of connected larger and denser fractal-like clusters. The time-resolved SAXS data provides evidence that protein unfolding occurs on the edges of the fractal-like clusters, resulting in a population of unfolded proteins in the inter-cluster space between clusters. Identifying the key stages of assembly in protein networks constructed from force labile proteins provides a greater understanding of the importance of protein unfolding in hierarchical biomechanics in vivo and creates future opportunities to develop bespoke biomaterials for novel biomedical applications. Cite as: Hughes et al. (2025) [Dataset] Capturing the Impact of Protein Unfolding on the Dynamic Assembly of Protein Networks. https://doi.org/10.5518/1632 Related publication: Hughes et al. Capturing the Impact of Protein Unfolding on the Dynamic Assembly of Protein Networks. Soft Matter. Accepted (2025) Contact: Matt Hughes (phymhug@leeds.ac.uk), Lorna Dougan (L.Dougan@leeds.ac.uk) 2. TERMS OF USE --------------- Copyright University of Leeds, 2025. Unless otherwise stated, this dataset is licensed under a Creative Commons Attribution 4.0 International Licence: https://creativecommons.org/licenses/by/4.0/. 3. PROJECT AND FUNDING INFORMATION ---------------------------------- Title: MESONET: Exploiting in situ protein unfolding to understand and control mesoscopic network formation Dates: Aug 22 - Aug 27 Funding organisation: Horizon Europe Guarantee Grant no.: EP/X023524/1 4. CONTENTS ----------- All data is split between 2 folders named Rheo Data, and SAXS Data which contain the raw data from experiments shown in the associated publication. Rheology Data contains a csv file of the BSA hydrogel gelation curves SAXS Data contains dat files of the scattering curves for illuminated BSA undergoing photochemcial crosslinking labelled with the latest gelation time at which they were acquired i.e. BSA_minus_1min is all the SAXS data acquired between 0 and 1 minutes of gelation time. 5. METHODS ---------- Experimental and analysis methods are outlined in the main manuscript and supplementary infomation.