{"id":1128,"date":"2022-07-07T01:37:30","date_gmt":"2022-07-07T01:37:30","guid":{"rendered":"https:\/\/sites.rutgers.edu\/shi-lab\/?post_type=tribe_events&p=1128"},"modified":"2022-09-01T18:48:49","modified_gmt":"2022-09-01T18:48:49","slug":"simon-scheuring-ccb-colloquium","status":"publish","type":"tribe_events","link":"https:\/\/sites.rutgers.edu\/shi-lab\/event\/simon-scheuring-ccb-colloquium\/","title":{"rendered":"CCB Colloquium – Professor Simon Scheuring, Weill Cornell Medicine,"},"content":{"rendered":"

\"Simon<\/strong><\/p>\n

\u201cBreaking Speed and Resolution Limitations of High-Speed AFM for Membrane Protein Structure-Function Analysis\u201d<\/strong><\/span><\/h1>\n

Simon Scheuring1,2,*
\n1 Weill Cornell Medicine, Department of Anesthesiology, 1300 York Avenue, New York, NY-10065, USA.
\n2 Weill Cornell Medicine, Department of Physiology and Biophysics, 1300 York Avenue, New York, NY-10065, USA<\/p>\n

High-speed atomic force microscopy (HS-AFM) is a powerful technique that provides dynamic movies of biomolecules at work [1]. We successfully used HS-AFM to take movies and determine dynamic parameters of membrane trafficking systems, transporters and channels.<\/p>\n

To break current temporal limitations to characterize molecular dynamics using HS-AFM, we developed HS-AFM line scanning (HS-AFM-LS) and HS-AFM height spectroscopy (HS-AFM-HS), methods whereby we scan the HS-AFM tip along a single scan line or keep it at a fixed position, respectively, and detect the motions of the molecules under the tip. This gives sub-nanometer spatial resolution combined with millisecond and microseconds temporal resolution of molecular fluctuations. HS-AFM-LS and HS-AFM-HS can be used in conjunction with HS-AFM imaging, thus giving access to a wide dynamic range [1]. This allowed us most recently to determine the single molecule kinetics of wild-type bacteriorhodopsin [2].<\/p>\n

To break current resolution limitations, we developed Localization AFM (LAFM). By applying localization image reconstruction algorithms to peak positions in high-speed AFM and conventional AFM data, we increase the resolution beyond the limits set by the tip radius and reach quasi-atomic resolution on soft protein surfaces in native and dynamic conditions. The LAFM method allows the calculation of high-resolution maps from either images of many molecules or many images of a single molecule acquired over time, opening new avenues for single molecule structural analysis [3].<\/p>\n

I will review these recent achievements, and present novel data and methods to integrate and strengthen HS-AFM as a powerful tool in structural biology and molecular biophysics. We reason that the progress of AFM must comprise two axes: First, making AFM\/HS-AFM a better tool. With the above-mentioned methods, we made significant progress in this axis over the last ~5 years. Second, making AFM\/HS-AFM data an integrated part of the structural biology \/ molecular biophysics toolbox. Such, we are developing now methods and data analysis and interpretation methods that interface with cryo-EM and X-ray crystallography, where AFM\/HS-AFM can contribute the urgently needed information about structure, dynamics and conformations under close-to-native conditions.<\/p>\n

References:<\/strong>
\n[1] Heath et al. Nature Communications, 2018, 9(1):4983, High-Speed AFM Height Spectroscopy (HS-AFM-HS): Microsecond dynamics of unlabeled biomolecules.
\n[2] Perrino et al., Nature Communications, 2021 12(7225), doi.org\/10.1038\/s41467-021-27580-2, Single molecule kinetics of bacteriorhodopsin by HS-AFM.
\n[3] Heath et al. Nature, 2021, 594(7863):385\u2013390, doi:10.1038\/s41586-021-03551-x, Localization Atomic Force Microscopy.<\/p>\n","protected":false},"excerpt":{"rendered":"

\u201cBreaking Speed and Resolution Limitations of High-Speed AFM for Membrane Protein Structure-Function Analysis\u201d Simon Scheuring1,2,* 1 Weill Cornell Medicine, Department of Anesthesiology, 1300 York Avenue, New York, NY-10065, USA. 2 … Read More<\/a><\/p>\n","protected":false},"author":790,"featured_media":0,"comment_status":"closed","ping_status":"closed","template":"","meta":{"_acf_changed":false,"_tribe_events_status":"","_tribe_events_status_reason":"","footnotes":""},"tags":[],"tribe_events_cat":[],"class_list":["post-1128","tribe_events","type-tribe_events","status-publish","hentry"],"acf":[],"_links":{"self":[{"href":"https:\/\/sites.rutgers.edu\/shi-lab\/wp-json\/wp\/v2\/tribe_events\/1128"}],"collection":[{"href":"https:\/\/sites.rutgers.edu\/shi-lab\/wp-json\/wp\/v2\/tribe_events"}],"about":[{"href":"https:\/\/sites.rutgers.edu\/shi-lab\/wp-json\/wp\/v2\/types\/tribe_events"}],"author":[{"embeddable":true,"href":"https:\/\/sites.rutgers.edu\/shi-lab\/wp-json\/wp\/v2\/users\/790"}],"replies":[{"embeddable":true,"href":"https:\/\/sites.rutgers.edu\/shi-lab\/wp-json\/wp\/v2\/comments?post=1128"}],"version-history":[{"count":2,"href":"https:\/\/sites.rutgers.edu\/shi-lab\/wp-json\/wp\/v2\/tribe_events\/1128\/revisions"}],"predecessor-version":[{"id":1171,"href":"https:\/\/sites.rutgers.edu\/shi-lab\/wp-json\/wp\/v2\/tribe_events\/1128\/revisions\/1171"}],"wp:attachment":[{"href":"https:\/\/sites.rutgers.edu\/shi-lab\/wp-json\/wp\/v2\/media?parent=1128"}],"wp:term":[{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/sites.rutgers.edu\/shi-lab\/wp-json\/wp\/v2\/tags?post=1128"},{"taxonomy":"tribe_events_cat","embeddable":true,"href":"https:\/\/sites.rutgers.edu\/shi-lab\/wp-json\/wp\/v2\/tribe_events_cat?post=1128"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}