Glycosyltransferases and more, important tools for drug design A. Bechthold, Universität Freiburg

1. Glycosyltransferases and more, important tools for drug design A. Bechthold, Universität Freiburg

4. Compounds with anticancer activity

5. Compounds with antimicrobial activity

7. 9 Sugars 6 GTs 6 Sugars 4 GTs 17 Sugars 10 GTs

8. UrdGT1a SaqGT5 SaqGT6 SaqGT1 SaqGT2 SaqGT3 UrdGT2 UrdGT1c SaqGT4 (?) UrdGT1b SaqGT3 SaqGT4 (?) LanGT1 LanGT3 LanGT2 LanGT4 LanGT1 LanGT4

9. Can we use glycosyltransferases as „tools“ to generate novel antibiotics?

10. LanGT1 LanGT3 LanGT1 LanGT2 LanGT4 LanGT4

11. LanGT1 LanGT3 LanGT4 UrdGT2

13. Landomycin A Urdamycin A More than 50 new compounds by combining landomycin- and urdamycin biosynthetic genes !

14. How do glycosyltransferases look like?

15. MurG GtfA OleD GtfB UrdGT2 OleI GtfD

16. UrdGT2

17. Can we combine N- and C-terminal parts of different glycosyltransferases?

19. +

20. LanGT1/LndGT1: 390 aa (25% non -identical aa)

21. LanGT1-like activity: Landomycinwith 5 and 6 sugars LndGT1-like activity: Landomycin with 2, 3 and 4 sugars no activity: Landomycin with 1 sugar

22. N-term. C-term.

23. Which amino acids are responsible for substrate specificity?

24. Mutations affecting the acceptor substrate specificity of a glycosyltransferase Hoffmeister et al., 2001, ChemBiol Hoffmeister et al., 2002, ChemBiol Hancock et al., 2006, Curr Opin Chem Biol Williams et al., 2007, Nat Chem Biol Williams et al., 2008, ChemBiol

25. α3 ß10 ß3 α10 α2 ß9 ß2 H4 H11 α9 α1 ß1 ß8 ß11 ß4 α5 α12 ß12 ß5 α8 αH13 α6b α6a ß13 ß6 α7 ß7 α16 α14 α15 v1a: β1-α3 v1b: region between α3 and α4 v2a: α4 v2b: β4-β5 v3: α6-α7 α4 N C

26. v1a v1b v2a v2b v3 N-term. C-term. v1a (64 aa): β1-α3 v1b (19 aa): region between α3 and α4 v2a (23 aa): α4 v2b (35 aa): β4-β5 v3 (68 aa): α6-α7

27. C

28. α3 ß10 ß3 α10 α2 ß9 ß2 H4 H11 α9 α1 ß1 ß8 ß11 ß4 α5 α12 ß12 ß5 α8 αH13 α6b α6a ß13 ß6 α7 ß7 α16 α14 α15 v1a: β1-α3 v1b: regionbetweenα3 andα4 v2a: α4 v2b: β4-β5 v3: α6-α7 α4 N C

30. α3 ß10 ß3 α10 α2 ß9 ß2 H4 H11 α9 α1 ß1 ß8 ß11 ß4 α5 α12 ß12 ß5 α8 αH13 α6b α6a ß13 ß6 α7 ß7 α16 α14 α15 v1a: β1-α3 v1b: regionbetweenα3 andα4 v2a: α4 v2b: β4-β5 v3: α6-α7 α4 N C

31. v1b I64 V67 A68 → V64 L67 A68 A73 E74 A75 → L73 Q74 S75 D82 → E82

32. v2b G107 R110 → R107 G110 L119 Y121 V122 F124 → I119 F121 I122 L124 T129 → D129

33. v2b G107 R110 → R107 G110 L119 Y121 V122 F124 → I119 F121 I122 L124 T129 → D129

34. v2a T89 → S89 R93 P94 D97 → P93 Q94 E97 E101 → A101 A104 → D104

35. I64 V67 A68 R93 P94 D97 L119 Y121 V122 F124 → V64 L67 G68 P93 Q94 E97 I119 F121 I122 L124

36. α3 ß10 ß3 α10 α2 ß9 ß2 H4 H11 α9 α1 ß1 ß8 ß11 ß4 α5 α12 ß12 ß5 α8 αH13 α6b α6a ß13 ß6 α7 ß7 α16 α14 α15 α4 N C

37. ERecognition α4 α3 β5 Positioning β4

38. Glycosyltransferases and more, important tools for drug design A. Bechthold, Universität Freiburg

39. Weitnauer et al., ChemBiol,2001 Weitnauer et al., Microbiol, 2002 Mosbacher et al., J Mol Biol 2003 Treede et al., Mol. Microbiol 2003 Weitnauer et al., ChemBiol 2004 Mosbacher et al., J Mol Biol, 2005 Treede et al., Ap Env Mic, 2005 Hofmann et al., ChemBiol, 2005 Boll et al., JBC, 2006

44. AviQ1 AviE2