1. 1 175.2

2. 2 Dual Picket Fence (K)L/D|I|L|D|Q|E|R(A) 175.2 113 113 115 128 129 129 128 115 113 113 115

3. 3

4. 4 Dual Picket Fence 113 71 99 99 115 99 115 99 99 71 113 (K)N D|L|A|V|V|D|V\R(I)

5. 5

6. 6 A E/D|T|A|L|Y|Y|C\A\K 115 101 71 113 163 163 163 163 113 71 101 115 Dual Picket Fence

7. 7

8. 8 Sparse Dominant Fragmentation 115 202 202 115 (K)I S R|P G D|S D|D|S R(S) Non-mobile proton z pre < #Arg

9. 9

10. 10 Dominant Cleavage Proline N-side N F|P/S/P V D/A A F R y9y9 b2b2 2887 97

11. 11

12. 12 Diagnose Doubly Charged Fragment Ions I/A|D|A|H|L|D|RI/A|D|A|H|L|D|R

13. 13

14. 14 Cry Babies (b-H 2 O & b pairs) E/H/A|V/E|G/D|C D|F Q L L K P(m/z)-H 2 O P(m/z)-2H 2 O

15. 15

16. 16 E/C|L/Q/T/C/RE/C|L/Q/T/C/R P(m/z)-H 2 O P(m/z)-2H 2 O 160 101 128 113 113 128 101 160 Cry Babies (b-H 2 O w/o b)

17. 17 733.5 942.4 1638.7 1043.6 1443.7 1691.4 1838.7

18. 18 Long & Extensive (R)T V T / V / W E / L / I | S|S/E/Y|F|T|A|E|Q R(Q) 733.5 942.4 1638.7 1043.6 1443.7 1691.4 1838.7

19. 19 709.4 594.4

20. 20 Dominant Ions – PM y++-h2o @ yn-2 DnEST 709.4 594.4 (K)V/A|E|I/E|H|A\E\K(E)

21. 21 599.4

22. 22 Dominant Ions – PM y++-h2o @ yn-2 DnEST (R)V L/H|T|A|L|H|S|S S S Y R(E) 599.4

23. 23 262.2 338.5 460.4 764.4

24. 24 262.2 338.5 460.4 764.4 Phosphopeptide Unambiguous Site (R)N Q|L|V|t/P E|P T / S R(A)

25. 25 429.4 659.6 699.3 544.3 560.2

26. Localizing a Phosphorylation Site Unambiguous 26 L/F|P/A/D|T/s/P/S T A\T K 429.4 659.6 699.3 544.3 560.2

27. 27 712.3 1511.2

28. Localizing a Phosphorylation Site Ambiguous 28 D S A|I|P/V/E|S D t D/D E|G\A\P R 712.3 1511.2 P(m/z) -H 3 PO 4 879

29. 29 1247.7 1293.5 1279.6 1210.4 545.4 598.7 452.4

30. 30 Phosphopeptide Ambiguous Site (R)L P/S s|P/V|Y/E/D|A|A/S|F K(A)

31. 31 343.3

32. 32 Met Oxidation y 6 -SOCH 4 y 8 -SOCH 4 y 9 -SOCH 4 (K)K/D|V|D|G A \ Y\m\T\K(V)

33. 33

34. 34 (R)G V D L D Q L/L|D|M|S|Y|E|Q|L|m|Q|L/Y S A R(Q) (R)G V D L D Q L/L|D|m|S/Y/E|Q|L|M|Q|L/Y S A R(Q) Met Oxidation – localizing the site

35. 35

36. 36 (R)q L/Q|L|A|Q|E|A|A\Q\K(R) (R)Q L/Q/L/A|Q/E/A|A Q\K(R) P(m/z)-NH 3 -17 Da Q to q Stinkers (b-NH 3 ) & Pyroglutamic Acid

37. 37

38. 38 (R)N P P R\F A\F|V|E|F|E|D|P R(D) (R)R G G P P\F A\F|V|E|F|E|D|P R(D)

39. 39 (R)N/P/P R F A|F|V|E|F|E|D|P R(D) (R)R G G P P\F|A|F|V|E|F|E|D|P R(D)