2018/12/17 by Hugo Sanabria, Sanabria, Hugo, Dmitro Rodnin +21
Biochemistry, Genetics and Molecular Biology · Materials Science · #Biomolecules (q-bio.BM) #Enzyme Structure and Function #FOS: Biological sciences #Photosynthetic Processes and Mechanisms #Protein Structure and Dynamics #Quantitative Methods (q-bio.QM)
paper · pdf · doi:10.48550/arxiv.1812.06937
openalex publication_date 2018/12/17 · openalex created_date 2022/08/01 · openalex updated_date 2026/07/28
We used a hybrid fluorescence spectroscopic toolkit to monitor T4 Lysozyme\n(T4L) in action. By unraveling the kinetic and dynamic interplay of the\nconformational states, we sought to elucidate the dynamic structural biology of\nT4L. In particular, by combining single-molecule and ensemble multiparameter\nfluorescence detection, EPR spectroscopy, mutagenesis, and FRET-positioning and\nscreening, we characterized three short-lived conformational states within the\nconformational landscape of the T4L over the ns-ms timescale. The use of 33\nFRET-derived distance sets, to screen known T4L structures, revealed that T4L\nin solution mainly adopts the known open and closed states in exchange at 4\n\μs. A newly found minor state, undisclosed by at present more than 500\ncrystal structures of T4L and sampled at 230 \μs, may be actively involved\nin the product release step in catalysis. The presented fluorescence\nspectroscopic toolkit is anticipated to accelerate the development of dynamic\nstructural biology.\n