551-1412-00L  Molecular and Structural Biology IV: Visualizing Macromolecules by X-Ray Crystallography and EM

SemesterSpring Semester 2019
LecturersN. Ban, D. Böhringer, T. Ishikawa, M. A. Leibundgut, K. Locher, M. Pilhofer, K. Wüthrich, further lecturers
Periodicityyearly recurring course
Language of instructionEnglish

AbstractThis course provides an in-depth discussion of two main methods to determine the 3D structures of macromolecules and complexes at high resolution: X-ray crystallography and cryo-electron microscopy. Both techniques result in electron density maps that are interpreted by atomic models.
ObjectiveStudents will obtain the theoretical background to understand structure determination techniques employed in X-ray crystallography and electron microscopy, including diffraction theory, crystal growth and analysis, reciprocal space calculations, interpretation of electron density, structure building and refinement as well as validation. The course will also provide an introduction into the use of cryo-electron tomography to visualize complex cellular substructures at sub-nanometer resolutions, effectively bridging the resolution gap between optical microscopy and single particle cryo-electron microscopy. Lectures will be complemented with practical sessions where students will have a chance to gain hands on experience with sample preparation, data processing and structure building and refinement.
ContentFebruary 22 Lecture 1 Prof. Dr. Kurt Wüthrich
History of Structural Molecular Biology

March 1 Lecture 2 Prof. Dr. Kaspar Locher
X-ray diffraction from macromolecular crystals

March 8 Lecture 3 Prof. Dr. Kaspar Locher
Data collection and statistics, phasing methods

March 15 Lecture 4 Prof. Dr. Nenad Ban
Crystal symmetry and space groups

March 22 Lecture 5 Ban Lab
Practical session with X-ray data processing

March 29 Lecture 6 Prof. Dr. Takashi Ishikawa
Principle of cryo-EM for biological macromolecules I, including hardware of TEM and detectors, image formation principle (phase contrast, spherical aberration, CTF), 3D reconstruction (central-section theorem, backprojection, missing information)

April 5 Lecture 7 Dr. Daniel Boehringer
Single particle analysis, including principle (projection matching, random conical tilt, angular reconstitution)

April 12 Lecture 8 Ban Lab
Practical session including specimen preparation (cryo, negative stain, visit to ScopeM

May 3 Lecture 9
Prof. Dr. M. Pilhofer
Tomography I, including basics and subtomogram averaging

May 10 Lecture 10 Ban Lab
Practical session with example initial EM data processing

May 17 Lecture 11 Prof. Dr. Martin Pilhofer
Practical session (including recent techniques, including cryo-FIB)

May 24 Lecture 12 Prof. Dr. Nenad Ban
EM and X-ray structure building, refinement, validation and interpretation

May 31 Lecture 13 Ban Lab
Practical session with model building and refinemen