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Protein structure determination using X-ray crystallography is the major experimental technique in Structural Biology. This module will provide an in-depth introduction to the technique. No similar module is offered at this level anywhere in South Africa.
Sub-modules
Lecturers
Programme Co-ordinator, Trevor Sewell, has experience in computer-based education, and is actively engaged in research in the field of structural biology. In addition to his co-ordinating role, Prof Sewell will present lectures on the determination of the three-dimensional structures of cells using tomography and single particle methods for the determination of macromolecular structure. He will also present workshops explaining some of the major achievements of structural biology studies on large structures including the ribosome.
Senior Lecturer, Muhammed Sayed (Department of Biotechnology, UWC), is a protein crystallographer who completed his post-doctoral studies under Professor Tom Blundell at Cambridge University. He will supervise the web-courses in protein structure and protein crystallography. His goal is to develop protein crystallography as a practical discipline. He will be involved in the “Introduction to X-ray crystallography” sub-module.
International consultant, Mike Lawrence, will also present lectures and workshops on protein crystallography
This module descriptor document also can be downloaded as an MS-Word document.
Main Outcomes
The ability to:
- crystallize proteins,
- collect and process crystallographic data,
- determine space groups,
- solve structures by molecular replacement,
- solve structures by isomorphous replacement and multiple anomalous dispersion techniques
- display structures
- build models of proteins using experimentally determined maps
- use standard crystallographic software
Main Content
- Basic theory of scattering
- Symmetry
- Design and operation of experimental equipment
- Data processing
- Structure determination
- Display and model building
- Concepts of structure refinement
Prerequisites
Heavy atom Derivatives
Read the sections on MIR and MAD in the courseware (Birkbeck Macromolecular Structure Determination Course):
- http://sbru.salamanderthemes.net/online_content/MSD-UCT/mir/index.htm
- http://sbru.salamanderthemes.net/online_content/MSD-UCT/mira/index.htm
- http://sbru.salamanderthemes.net/online_content/MSD-UCT/mad/index.htm
- http://sbru.salamanderthemes.net/online_content/MSD-UCT/mada/index.htm
Processing
- Process the psaa PtNit derivative data using either DENZO of CrystalClear
Map Interpretation
- install the latest Windows version of O.
- Do the O tutorial (http://sbru.salamanderthemes.net/online_content/MSD-UCT/modela/index.htm)
- read about the theory of electron density modelling. (http://sbru.salamanderthemes.net/online_content/MSD-UCT/model/index.htm)
- You can use the PSAA maps from
\\rotweiler\download\psaa
Programme details
- Read the manuals for O, Solve, Resolve, MAID.
Online Lectures
Birkbeck Protein Crystallisation Course
http://sbru.salamanderthemes.net/online_content/birkbeck_protein_crystallisation/index.htm
Birkbeck Macromolecular Structure Determination Course
Web course located at
http://sbru.salamanderthemes.net/online_content/MSD-UCT/xtalindx.htm
Electron Crystallography Virtual Course – Erice 2004
Protein Crystallography Practical Course
Gwen Nneji
- Lecture 1: Basic Principles of Protein Crystallography
- Lecture 2: Strategies, Optimisations, Non-Standard Methods
Tutorials
[PC] Tutorial 1: X-ray data processing by Mike Lawrence
[PC] Tutorial 2: Phasing by isomorphous replacement by Mike Lawrence
References
[bibliplug last_name=”blow” category=”PC”]
[bibliplug last_name=”lawrence” category=”PC”]
Manuals
Solve/Resolve – see the software manuals section for Solve/Resolve and other software, such as PyMol, O, Spider, etc.