Cryo-Electron Microscopy in Structural Biology From Structural Insights to Tomography and Drug Discovery

SECTION I: Cryo-EM in Cell Biology - Structure Determination of Cellular Organelle 1. Atomic-resolution protein structure determination by cryo-EM 2. Sub-ångström cryo-EM Structure of a prion proto-fibrils 3. Cryo-EM structure of native human uromodulin, a zona pellucida module polymer 4. Chromatosome, telomere structure and genome maintenance using cryo-EM 5. Cryo-EM of the dynamin super-family proteins 6. Mito-ribosome structure using cryo-EM SECTION II: Technology Platform & Evolving Technologies 1. Cryo-EM data in the cloud computing 2. The silent revolution of cryo-EM and GPCR structures 3. Development of world-first multi-gas plasma focused ion beam microscope 4. 3-D printed cell culture grid holders for improved cellular specimen preparation in cryo- electron microscopy 5. Local resolution estimates of cryo-EM reconstruction 6. A new cryo-EM system for electron 3D crystallography SECTION III: Cryo-EM in Virology - High-Resolution Structures of Viruses 1. Cryo-Electron Microscopy structure of adeno-associated virus-DJ bound by a heparinoid pentasaccharide 2. Cryo-EM and tomography of vesicular stomatitis viruses 3. High-resolution structures of HIV-1 gag cleavage mutants 4. Cryo-electron microscopy of the T7 bacteriophage portal and fiber-less tail complex 5. SARS-Co-2 spike protein structure determination SECTION IV: Cryo EM in Membrane Biology – Probing the structures of Membrane Proteins 1. Structure and activity of lipid bilayer within a membrane-protein transporter 2. Cryo-EM structure of human Rhodopsin bound to an inhibitory G protein 3. Cryo-EM structure of the acetylcholine receptor that cause congenital myasthenic syndromes 4. Structure of the mitochondrial F-ATPase as determined by electron cryo-microscopy 5. Cryo-EM structure of cardiac amyloid fibrils from an immunoglobulin light chain AL amyloidosis patient SECTION V: Cryo-EM in Molecular Biology - Structural Insights of Nuclear and Gene Machineries 1. Structural information from cryo-EM studies on multicomponent molecular assemblies such as those found in DNA-double-strand-break repair 2. Cryo-EM structure of the DNA-PK holoenzyme 3. Cryo-EM reconstruction at 4.0 Å of human Polymerase III 4. Cryo-electron microscopy structures of UV-DDB bound to nucleosome 5. Cryo-electron microscopy of the chromatin fiber 6. Time-resolved cryo-EM studies in the ribosome dynamics SECTION VI: Cryo-Tomography and Emerging Technologies 1. Cryo-Electron Tomography in membrane protein structural research 2. Cryo-tomography of membrane transporters involved in neurological psychiatric disease 3. Enveloped viruses and coated vesicles - electron cryo-microscopy and tomography 4. Electron cryo-tomography in the study of autophagy or bacterial secretion systems 5. DNA Origami sign posts for identifying proteins on cell membranes by cryo-tomography 6. 3-D deconvolution processing for STEM cryo-tomography SECTION VII: Applications of Cryo-EM/ET in Drug Discovery 1. Cryo-EM to enable research and drug discovery of CD20 in complex with the therapeutic monoclonal antibody rituximab 2. Fragment-based drug discovery using cryo-EM 3. CryoEM approaches to study allosteric regulation of GTP cyclohydrolase I 4. Cryo-EM in drug discovery 5. Structural insights of an advanced vaccine candidate as derived from SARS-CoV-2 spike protein 6. Unleashing the power of Cryo-EM Epilogue

Biography

Krishnarao Appasani trained as a molecular cell biologist, receiving Masters and Doctoral degrees in Biochemistry and Molecular biology. He accepted a Lectureship at Boston University and taught cell and molecular biology courses to undergraduates and graduate students. He was a post-doc in the laboratory of Nobel Laureate, the late Prof. H. Gobind Khorana at the Massachusetts Institute of Technology. Subsequently, he joined faculty of Harvard Medical School and directed the Thoracic Oncology Laboratory at the Brigham &Women’s Hospital and the Dana-Farber Cancer Institute in Boston. During this time he completed an MBA at Bryant University, Smithfield, RI. And then moved Perkin Elmer and directed the Microarrays group. Dr Appasani has published about 60 research papers and abstracts in international journals, especially on Gene Expression. He was awarded the “outstanding investigator award” by the American Federation for Medical Research, Washington D.C., competed globally for the Ranbaxy Research Medal, and was honored with the “1997-Ranbaxy Research Award in the field of Applied Medical Sciences.” At Perkin Elmer, Dr Appasani organized a global Biomics Seminar series to educate scientists and to initiate & develop collaborations with researchers in academia and other industries. For about 15 years he ran multi-million-dollar knowledge management enterprise and served as an international authority in bridging Academia and Industry by fostering inter-disciplinary and Translational Science. In 2016 Dr Appasani was inducted as Life Fellow of Royal Society of Biology, England for his global efforts in promoting science education.