Back

Conformationally Adaptive Dyes Enable Protein-Adaptive Differential Scanning Fluorometry

Wu, T. S.; Yu, J. C.; Suresh, A.; Gale-Day, Z. J.; Alteen, M. G.; Woo, A. S.; Millbern, Z.; Johnson, O. T.; Carroll, E. C.; Partch, C. L.; Fourches, D.; Vilnueza, N. R.; Vocadlo, D. J.; Gestwicki, J. E.

2023-01-24 biochemistry
10.1101/2023.01.23.525251 bioRxiv
Show abstract

Flexible in vitro methods alter the course of biological discoveries. Differential Scanning Fluorimetry (DSF) is a particularly versatile technique which reports protein thermal unfolding via fluorogenic dye. However, applications of DSF are limited by widespread protein incompatibilities with the available DSF dyes. Here, we enable DSF applications for 66 of 70 tested proteins (94%) including 10 from the SARS-CoV2 virus using a chemically diverse dye library, Aurora, to identify compatible dye-protein pairs in high throughput. We find that this protein-adaptive DSF platform (paDSF) not only triples the previous protein compatibility, but also fundamentally extends the processes observable by DSF, including interdomain allostery in O-GlcNAc Transferase (OGT). paDSF enables routine measurement of protein stability, dynamics, and ligand binding. One-Sentence SummaryNext generation protein-adaptive DSF (paDSF) enables rapid and general measurements of protein stability and dynamics.

Matching journals

The top 5 journals account for 50% of the predicted probability mass.

50% of probability mass above

"Similar papers" are the closest papers from that journal in the model's embedding space. They show what the match is built on, but the ranking comes mostly from a classifier over the whole training set, not from these examples alone.