1. Małopolska Centre of Biotechnology (MCB) X-Ray Crystallography Laboratory Looking into the deep – structural investigations of biological macromolecules

2. X-Ray Crystallography Laboratory Head: Grzegorz Dubin, PhD 5 post-doctoral researchers 7 Ph.D. students Research methodology and specialized equipment: The X-ray Crystallography Laboratory of MCB has extensive experience in solving protein structures and the structures of complexes. The primary laboratory equipment consists of an X-ray diffractometer. Crystallization screening is carried out using an automatic pipetting station. Laboratory is also exceptionally well equipped for expression, purification and analysis of proteins. Research themes We are primarily interesting study in interactions of proteins and proteins and low molecular weight substances within the subject of carcinogenesis and bacterial pathogenesis. Currently were realize our own projects involving structural characterization of kinases of essential role cancer, p53 pathway, bacterial proteases of unknown catalytic mechanism and staphylococcal proteases. In parallel to the crystallographic studies we also develop aptamer selection techniques (especially single- stranded DNA) against targets of importance in cancer.

3. Contents of the cell water, small molecules, lipids, nucleic acids, proteins … Proteins are the bodymakers, involved in many cellular processes and/or diseases. How can we see the protein?

4. Production of macromolecules METHODS genetic engineering molecular biology chromatography biochemistry Proteins in appropriate chemical environment may form crystals Gene encoding protein of intrest (e.g. protein of human origin) Bacterium - protein production „factory” Protein in a test tube

5. In order to measure something accurately, you need the appropriate ruler…. When using light to measure an object, the wavelength of the light needs to be similar to the size of the object. X-rays, with wavelengths of approximately 0.5 to 1.5 angstroms (Å)*, can measure the distance between atoms. *1 Å = 10 -10 m Looking at the atomic scale

6. X-ray crystallography reveals protein structure Synchrotron X-ray source Crystallography produced the largest number of Nobel Laureates throughout history... …28 Nobel Prizes Cartoon representation of protein structures

7. Take home message Proteins are involved in most of the physiological processes X-ray crystallogaphy is the way to get information about protein structure Knowledge of protein structure may be used in structure based drug design – producing new chemical compounds which may serve as drugs