Journal article
Global analysis of endogenous protein disorder in cells
S Zhang, TC Owyong, O Sanislav, L Englmaier, X Sui, G Wang, DW Greening, NA Williamson, A Villunger, JM White, B Heras, WWH Wong, PR Fisher, Y Hong
Nature Methods | Published : 2025
Abstract
Disorder and flexibility in protein structures are essential for biological function but can also contribute to diseases, such as neurodegenerative disorders. However, characterizing protein folding on a proteome-wide scale within biological matrices remains challenging. Here we present a method using a bifunctional chemical probe, named TME, to capture in situ, enrich and quantify endogenous protein disorder in cells. TME exhibits a fluorescence turn-on effect upon selective conjugation with proteins with free cysteines in surface-exposed and flexible environments—a distinctive signature of protein disorder. Using an affinity-based proteomic approach, we identify both basal disordered prote..
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Awarded by La Trobe University
Funding Acknowledgements
This study was funded by grants to Y.H. from the Australian Research Council (FT210100271 and DE170100058), the National Health and Medical Research Council (APP1161803 and APP2029017) and the Rebecca L. Cooper Medical Research Foundation (PG2018043). S.Z. was supported by a La Trobe University Graduate Research Scholarship, a La Trobe University Full Fee Research Scholarship and an LIMS Writing-up Award. We thank the Bio21 Mass Spectrometry and Proteomics Facility and La Trobe University Bioimaging and Comprehensive Proteomics Platform for the professional support and access to the equipment. We thank the assistance of resources and services from the National Computational Infrastructure by the Australian government. This research was undertaken in part using the MX1 beamline and MX2 beamline (using the Australian Cancer Research Foundation detector) at the Australian Synchrotron, part of ANSTO. We are grateful to D. Loesch (La Trobe University), A. Atkinson (La Trobe University), A. Evans (Royal Melbourne Hospital) and E. Storey (Monash University/Alfred Hospital) for assistance with clinical information and original participant recruitment. We also thank D. M. Hatters (University of Melbourne), M. Lee (La Trobe University), T. Pukala (University of Adelaide), M.-C. Giel (La Trobe University) and R. Xu (La Trobe University) for helping with sample preparation, sharing original plasmids and helpful discussions, and C. Allan (La Trobe University) for isolation of the original lymphoblastoid cell lines.