PT  - JOURNAL ARTICLE
AU  - Megan Leander
AU  - Zhuang Liu
AU  - Qiang Cui
AU  - Srivatsan Raman
TI  - Deep mutational scanning and machine learning reveal structural and molecular rules governing allosteric hotspots in homologous proteins
AID  - 10.1101/2022.05.01.490188
DP  - 2022 Jan 01
TA  - bioRxiv
PG  - 2022.05.01.490188
4099  - http://biorxiv.org/content/early/2022/05/01/2022.05.01.490188.short
4100  - http://biorxiv.org/content/early/2022/05/01/2022.05.01.490188.full
AB  - A fundamental question in protein science is where allosteric hotspots – residues critical for allosteric signaling – are located, and what properties differentiate them. We carried out deep mutational scanning (DMS) of four homologous bacterial allosteric transcription factors (aTF) to identify hotspots and built a machine learning model with this data to glean the structural and molecular properties of allosteric hotspots. We found hotspots to be distributed protein-wide rather than being restricted to “pathways” linking allosteric and active sites as is commonly assumed. Despite structural homology, the location of hotspots was not superimposable across the aTFs. However, common signatures emerged when comparing hotspots coincident with long-range interactions, suggesting that the allosteric mechanism is conserved among the homologs despite differences in molecular details. Machine learning with our large DMS datasets revealed that global structural and dynamic properties to be a strong predictor of whether a residue is a hotspot than local and physicochemical properties. Furthermore, a model trained on one protein can predict hotspots in a homolog. In summary, the overall allosteric mechanism is embedded in the structural fold of the aTF family, but the finer, molecular details are sequence-specific.Competing Interest StatementThe authors have declared no competing interest.