1. PRESENTATION II NAME By Christine Harvey Dihydrofolate Reductase Beta lactmase p300 Macrophage Infectivity Potentiator

2. Dihydrofolate Reductase in Escherichia coli

3. http://en.wikipedia.org/wiki/DHFR Class: oxidoreductase Reduces dihydrofolate (DHF) to tetrahydrofolate (THF) THF is a precursor to amino acids, purine and thymidylate. DHFR is imperative to cell life

4. Function Source: Schweitzer, Barry I., et al. The FASEB Journal Vol 4, “Dihydrofolate reductase as a therapeutic target.” May 1990.

5. Mechanism http://en.wikipedia.org/wiki/DHFR

6. Structure

7. Binding Pocket

9. MOVIE

14. Sequence Alignment

15. Phylogenetic Tree

16. Beta-Lactamase in Escherichia coli

17. Class: Hydrolase Beta Lactamase is produce by bacteria after being exposed to beta lactam antibiotics Responsible for beta lactam antibiotic resistance

18. Penicillin Contains beta lactam ring

19. Mechanism

21. 21

28. p300 In Homo Sapiens

29. Class: Transferase p300 transfers an acetyl group from acetyl coenzyme A to an active site lysine Transcriptional activation occurs upon acetyl transfer

30. Title p300 is responsible for cell growth before and after birth Cancer suppressor

31. Domains Taz2 KIX Bromodomain HAT Zinc Fingers (3) Taz2 Creb

32. Histone Acetyl Transferase Active site tyrosine acts as nucleophile Source: http://www.bmolchem.wisc.edu/labs/denu/images/hats/sequential.jpghttp://www.bmolchem.wisc.edu/labs/denu/images/hats/sequential.jpg

37. Docking

39. 39

40. Macrophage Infectivity Potentiator in Legionella pneumophila

41. Legionella pneumophila gram negative bacteria Causes Legionnaires’ disease Carried in aerosols from man-made water systems

42. MIP Class: Isomerase Catalyzes the isomeration of the N- terminal on proline Cause of virulence in Legionnaires’ disease Located on outer membrane protein 25 kDa

43. Crystalization Solution: 100 mM MES (pH 6.1–6.5), 15–20% (w/v) PEG 8000 and 500 mM zinc acetate. Protein concentration was 10 mg ml–1in 20 mM HEPES, pH 7.0 Crystals were grown for 2-3 weeks at 288K X-ray data was collected at 100K in a nitrogen gas stream (dried paraffin oil) at 2.4 Angstrom resolution Multiwavelength anomalous dispersion (MAD) with zinc was used for phasing

44. 44 Source: Riboldi-Tunnicliffe, Alan et al., Nature Structural Biology, vol.8: 9. “Crystal structure of Mip, a prolylisomerase from Legionella pneumophila.” September 2001.

45. Mechanism Peptidyl-prolyl cis-trans isomerases’ (PPIases) mechanism is not currently know Source: Park, Steven T., et al. Journal of Biological Chemistry. Vol. 267 No.5, “PPIase Catalysis by Human Fk506-binding Protein Proceeds Through a Conformational Twist Mechanism.” February 1992.

46. Source: Riboldi-Tunnicliffe, Alan et al., Nature Structural Biology, vol.8: 9. “Crystal structure of Mip, a prolylisomerase from Legionella pneumophila.” September 2001.

49. 49

51. 51

52. 52

53. 53 Questions?