1. By: Dario Marotta 10/23/2014 1 ELASTASE A SERINE PROTEASE

2. 1. Proteases = Protein  Trypsin  Chymotrypsin  Elastase 2. Pancreatic Lipase = Fat 3. Amylase = Carbohydrates 10/23/2014 2 ANATOMY

3. Composition  Hydrophobic Amino Acids  Glycine, Proline, Alanine, Valine Elastic Properties  Arteries  Lungs  Skin  Ligaments  Bladder 10/23/2014 3 ELASTIN

4. Step 1: Mechaelis Complex  Ser ~195 Hydroxyl Group = Nucleophile  Asp ~119 H-bond to His~57  His ~57 better (proton acceptor) Step 2: Tetrahedral Intermediate  His ~57 Accepts Ser ~195 Hydrogen Step 3: Deacylation  Water = nucleophile  Regenerates the catalytic triad 10/23/2014 4 REACTION MECHANISM

5. 10/23/2014 5 SERINE PROTEASES Specificity Determines Function Serine Protease specificity is determined by size of binding pocket

6. DeLano, W.L. 2002 The PyMOL Molecular Graphics System San Carlos, Ca, USA DeLano Scientific Valine ~190/~216  Isopropyl side chain restricts access  Alanine, Glycine, Valine 10/23/2014 6 SPECIFICITY Elastase Binding Pocket  Red = Catalytic Triad  Green = Valine ~190  Grey = Valine ~216

7. 7 SPECIFICITY

8. 8 ACTIVE SITE Catalytic triad Specificity residues block large side chains

9. 9 ACTIVE SITE Tetrahedral intermediate - ready to leave and regenerate binding pocket residues

10. Generally not a membrane-bound enzyme, although membrane-bound forms do exist. 10 HYDROPATHY PLOT

11. 10/23/2014 11 SEQUENCE ALIGNMENT FEBS Journal Volume 277, Issue 10, pages 2238-2254, 26 APR 2010 DOI: 10.1111/j.1742-4658.2010.07659.x http://onlinelibrary.wiley.com/doi/10.1111/j.1742-4658.2010.07659.x/full#f2 Chymotrypsin-like Serine Proteases  hPR3 ( Human proteinase 3 )  hNE ( Human neutrophil elastase ) Signal peptides: Blue N-terminal peptides: Green C-terminal peptides: Orange Catalytic Triad: Green Stars Glycosylation Sites: Orange Stars A-Helices: Yellow B-Strands: Purple

12. 10/23/2014 12 ADDITIONAL ORGANISMS

13. 13 ELASTASE 3B SEQUENCE ALIGNMENT (VARIOUS SPECIES) Conserved Amino Acids His 57, Asp 102, Val 190, Ser 195

14. 10/23/2014 14 HUMAN ELASTASE VARIATION

15. His 57, Asp 102, Val 190, Val 216 Ser 195 15 HUMAN ELASTASE SEQUENCE ALIGNMENT Conserved Amino Acids

16. 10/23/2014 16 STRUCTURE MAP FEBS Journal Volume 277, Issue 10, pages 2238-2254, 26 APR 2010 DOI: 10.1111/j.1742-4658.2010.07659.x http://onlinelibrary.wiley.com/doi/10.1111/j.1742-4658.2010.07659.x/full#f2 Catalytic Triad: Green Glycosylation Sites: Orange A-Helices: Yellow B-Strands: Purple

17. Emphysema Cystic Fibrosis Inflammation Coronary Artery Disease Liver Disease Pancreatitis 10/23/2014 17 CLINICAL SIGNIFICANCE