1. Reactant Product Energy Landscape

2. EcoRV

3. Phosphodiester hydrolysis H2OH2O

4. “Reactant State” Crystal Structure Cleavage Site Water Oxygen 3.7Å Ala 92 (Mutated from Lys)

5. Molecular Dynamics of Wild Type Lys 92 Water Molecule PETRA IMHOF

6. MD I Molecular Dynamics of Wild Type PETRA IMHOF

7. MD II

8. Principal Components I Principal Component

9. Molecular Dynamics of Wild-Type Deprotonated Lys 92 Water Molecule

10. Muscle Contraction Thick filamentThin filament

11. ATP Hydrolysis by Myosin SONJA SCHWARZL

12. Serine Protease Melanie Böttcher

13. Index Definition Digestive enzymes Blood clotting Complement system

14. Serine Protease Cutting of certain peptide bonds in other proteins Activity depends on a set of amino acid residues in the active site of the enzyme Based on nucleophilic attack of the targeted peptidic bond by a serine

15. Serine Amino acid alcohol as residue

16. Digestive enzymes Chymotrypsin, Trypsin, Elastase closely-similar structures different substrate specificities Pancreatic proteases  proenzymes trypsinogen and chymotrypsinogen  synthesized in the pancreas and secreted into the lumen of the small intestine

17. Mechanism

18. Active site

19. Trypsinogen-Trypsin loss of six amino acids from one end overall structures remain similar Ca++ for thermal stability activated enzyme does have more of its structure organized into sheets

20. Trypsin Trypsin cleaves peptide bonds on the C-terminal side of arginines and lysines  Alcaline amino acids Activation by enteropeptidase or through protoelyse Optimum pH 7 to 8

21. Chymotrypsinogen-Chymotrypsin Chain is clipped in four places  release of 2 dipeptides  creation of two breaks in the backbone 1-13 segment retained as part of the active enzyme, linked on by a disulfide bond Activation by trypsin

22. Chymotrypsin Active site: His57-Ser195-Asp102 Chymotrypsin cuts on the C- terminal side of tyrosine, phenylalanine, and tryptophan residues hydrolysis of bonds of leucyl, methionyl, asparaginyl and glutamyl residues.

24. Elastase cuts peptide bonds next to small, uncharged side chains such as those of alanine, serine, valine and threonin cuts collagen

25. Blood Clotting System Cascade of several mechanism  Involved Serine Proteases  Thrombin  Plasmin  Factor 10 a  Factor 11 b

27. Human Thrombin with the Amino acids 55-65 of Hirudin Thrombin Inactivated Prothrombin cleaves into activated Thrombin Thrombins cuts Paraglobuline (= fibrinoplastic substance) and forms the soluble state of it Precipitation by plasmasalt in unsoluble state

28. Serpins Serine Protease Inhibitors inhibit the action of their respective serine protease  serine protease binds the serpin instead of its normal substrate  protease makes a cut in the serpin leading to –the formation of a covalent bond linking the two molecules –a massive allosteric change in the tertiary structure of the serpin which moves the attached protease to a site where it can be destroyed

29. Serine ProteaseSerpin Chymotrypsinalpha-1-antichymotrypsin Complement factor C1s C1 Inhibitor (C1INH) Elastase (secreted by neutrophils)alpha-1-antitrypsin Clotting factor 10 (X)antithrombin III Thrombinantithrombin III Plasminalpha-2-antiplasmin Trypsinpancreatic trypsin inhibitor

30. Occurrence/ Function stop proteolytic activity blood plasma clotting complement systems where a tiny initial activating event leads to a rapidly amplifying cascade of activity

31. Serpin Deficiencies inherited diseases mutation in the encoding gene for serpin examples

32. Alpha-1-Antitrypsin Serpin for Elastase secreted by neutrophils (lung infection)  emphysema  liver damage tests are running

33. The Others Other Serine Proteases –Subtilisin –Acetylcholinesterase Serpinlike Molecules –Angiotensinogen –Chicken Ovalbumin