96047 - Crystallography and Cryo-Election Microscopy for Protein Structure Determination

Learning outcomes

At the end of the module, the student acquires the basic knowledge necessary for the determination of the structure of biological macromolecules at atomic level, using the most current structural techniques such as X-ray crystallography and cryo-electron microscopy. The student has developped the skills for a critical evaluation of the use of these techniques and to interpret the results of a structural investigation of biological macromolecules using crystallography or Cryo-EM.

Course contents

• Introduction to structural biology and biocrystallography;
• Crystallisation of biological macromolecules: basic principles of crystallisation, properties of crystals of biological macromolecules and main techniques for crystallisation of biological macromolecules;
• Crystal symmetry of biological macromolecules;
• X-ray diffraction: physical principles and Bragg's law;
• X-ray sources: conventional generators and synchrotron;
• Phase problem in crystallography and methods for structural resolution of proteins;
• Structure validation and deposit in the Protein Data Bank;
• Examples of crystal protein structures;
• Basic principles of electron microscopy (EM);
• Comparison of X-ray crystallography and cryo-EM in the determination of structures of biological macromolecules;
• Electron microscope;
• Sample preparation and data collection;
• Image analysis and structural resolution;
• Examples of cryo-EM structures of biological macromolecules.
  

Readings/Bibliography

Lecture slides and teaching material will be made available in the Virtual platform. Additional material such as scientific articles, reviews, films etc. useful for further study and exam preparation will be made available in Virtual.

Recommended texts for further study:

- B. Rupp «BIOMOLECULAR CRYSTALLOGRAPHY: Principles, Practice, and Application to Structural Biology” Kindle Edition, 2009.

- A. McPherson "Introduction to macromolecular crystallography" J. Wiley 2003 §

- D. Blow «Outline of Crystallography for Biologists», Oxford University Press 2002

- G. Rhodes "Crystallography made crystal clear" III ed., Academic Press 2006

Teaching methods

Lectures are face-to-face with slide shows.

Assessment methods

The learning assessment is conducted through an oral examination consisting of a presentation of a scientific article relevant to the course topics and subsequent questions on the course programme. The examination is designed to test the skills and abilities acquired during the course, including the ability to read in depth and present a scientific article in a comprehensive manner.

The time required for the oral examination is approximately 30 minutes.

Not sufficient evaluation: deficient knowledge of the course topics, errors in basic chemical and/or biological concepts, inappropriate language.

Sufficient evaluation: minimal knowledge of the topics covered in the course, poor argumentation skills, some even serious errors and barely appropriate language.

Good evaluation: good knowledge of the topics covered in the course and good argumentative and expository skills with correct language. Some errors and/or imperfections not serious.

Excellent evaluation: very good mastery, ability to elaborate and expound on the topics of the course. Use of highly appropriate language.

Students have the right to reject the proposed positive grade once (in accordance with the University Teaching Regulations, ART.16, paragraph 5).

Students with learning disorders and\or temporary or permanent disabilities: please, contact the office responsible (https://site.unibo.it/studenti-con-disabilita-e-dsa/en/for-students) as soon as possible so that they can propose acceptable adjustments. The request for adaptation must be submitted in advance (15 days before the exam date) to the lecturer, who will assess the appropriateness of the adjustments, taking into account the teaching objectives.

Students recognized as “working students”: please consult the dedicated website (https://www.unibo.it/en/study/guide-to-choosing-your-programme/balancing-study-and-work ) to apply for this status and to learn about the available measures.

Teaching tools

Videoprojector and PC. Power point presentations used during the lessons and any research material will be provided to the students in electronic format.

Office hours

See the website of Simona Fermani