Biophysical Society Thematic Meeting| Lima 2019

Revisiting the Central Dogma of Molecular Biology at the Single-Molecule Level

Thursday Speaker Abstracts

HIGH-SPEED ATOMIC FORCE MICROSCOPY: STRUCTURAL DYNAMICS OF SINGLE UNLABELED PROTEINS Simon Scheuring 1,2 ; 1 Weill Cornell Medicine, Anesthesiology, New York, NY, USA 2 Weill Cornell Medicine, Biophysics & Physiology, New York, NY, USA The advent of high-speed atomic force microscopy (HS-AFM(1)) has opened a novel research field for the dynamic analysis of single bio-molecules: Molecular motor dynamics (2,3), membrane protein diffusion (4), assembly (5) and conformational changes (6) could be directly visualized. Further developments for buffer exchange (7) and temperature control (8) during HS- AFM operation provide breakthroughs towards the performance of dynamic structural biochemistry using HS-AFM. I will exemplify the power of HS-AFM for the quantitative analysis of function-related structural dynamics on the membrane deformation complex ESCRT- III (5), a glutamate transporter homologue (6), and ligand-gated ion channels (9,10). Finally, I will introduce high-speed AFM line scanning (HS-AFM-LS) and high-speed AFM height spectroscopy (HS-AFM-HS) that reache millisecond and microsecond temporal resolution, respectively, of single molecule dynamics (11). References:1) Ando, Chem Rev 2014, 114(6):3120-882) Kodera, Nature 2010, 468(7320):72-63) Uchihashi, Science 2011, 333(6043):755-84) Casuso, Nat Nanotechnol 2012, 7(8):525-95) Chiaruttini, Cell 2015, 163(4):866-79.6) Ruan, PNAS 2017, 114(7):1584-15887) Miyagi, Nat Nanotechnol 2016, 11(9):783-908) Takahashi, Small 2016, 12(44):6106-61139) Ruan, 2018 115(41):10333- 1033810) Marchesi, Nature Communications, 2018, 9(1):397811) Heath and Scheuring, Nature Communications, 2018, 9(1):4983

13