Inhibitors of Serine Proteases from a Microcystis sp. Bloom ... - MDPI

Inhibitors of Serine Proteases from a Microcystis sp. Bloom-Material collected from Timurim Reservoir, Israel Rawan Hasan-Amer and Shmuel Carmeli* Raymond and Beverly Sackler School of Chemistry, Raymond and Beverly Sackler Faculty of Exact Sciences, Tel-Aviv University, Ramat Aviv, Tel-Aviv 69978, Israel.

Supplementary Material

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Table of content S1. Figure S1. Structures of the Metabolites and Isolation Artifacts Isolated from the Extract of IL-428 S2. Table S1. 1H NMR Data of 1 and its isolation artifacts 3-8 in DMSO-d6 S3. Table S2. 13C NMR Data of 1 and its isolation artifacts 3-8 in DMSO-d6 S4. 1H NMR Spectrum of Micropeptin TR1058 (1) in DMSO-d6 S5. 13C NMR Spectrum of Micropeptin TR1058 (1) in DMSO-d6 S6. HSQC Spectrum Micropeptin TR1058 (1) in DMSO-d6 S7. HMBC Spectrum of Micropeptin TR1058 (1) in DMSO-d6 S8. COSY Spectrum of Micropeptin TR1058 (1) in DMSO-d6 S9. TOCSY Spectrum of Micropeptin TR1058 (1) in DMSO-d6 S10. ROESY Spectrum of Micropeptin TR1058 (1) in DMSO-d6 S11. HR ESI MS data of Micropeptin TR1058 (1) S12. 1H NMR Spectrum of Aeruginosin TR642 (2) in DMSO-d6 S13. 13C NMR Spectrum of Aeruginosin TR642 (2) in DMSO-d6 S14. HSQC Spectrum of Aeruginosin TR642 (2) in DMSO-d6 S15. HMBC Spectrum of Aeruginosin TR642 (2) in DMSO-d6 S16. COSY Spectrum of Aeruginosin TR642 (2) in DMSO-d6 S17. TOCSY Spectrum of Aeruginosin TR642 (2) in DMSO-d6 S18. ROESY Spectrum of Aeruginosin TR642 (2) in DMSO-d6 S19. HR ESI MS data of Aeruginosin TR642 (2)

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S1. Figure S1. Structures of the Metabolites and Isolation Artifacts Isolated from the Extract of IL-428

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S2. Table S1. 1H NMR Data of 1 and its isolation artifacts 3-8 in DMSO-d6 Position Val

2 3 4 5 NH N-MeTyr 2 3a 3b 5,5' 6,6' OH NMe Leu 2 3a 3b 4 5 6 Ahp/Amp 3 4a 4b 5a 5b 6 NH OMe OH 2 Gln/Glu/ 3a 3b OMe Glu 4a 4b NH

1 4.73 m 2.06 m 0.73 d 0.85 d 7.53 d 4.91 m 2.66 d 3.09 d 6.88 d 6.62 d 9.20 s 2.70 s 4.59 dd 10.0,4.5 0.41 dt 10.0,1.5 1.54 dt 10.0,1.5 0.96 m 0.49 d 0.68 d 4.37 ddd 1.71 m 2.54 m 1.71 m 1.71 m 4.88 brs 7.34 d ---6.02 d 4.29 ddd 1.66 m 2.18 m 2.02 m 2.10 m 8.49 d

3 4.75 m 2.07 m 0.79 d 0.89 d 7.53 d 4.97 m 2.66 d 3.10 d 6.89 d 6.64 d 9.14 s 2.72 s 4.64 dd 0.31 dt 1.62 dt 0.99 m 0.49 d 0.70 d 4.41 1.72 ddd m 2.35 m 1.60 m 2.04 m 4.41 s 7.34 d 3.02 s ---4.31 m 1.65 m 2.19 m 2.11 m 2.02 m 8.55 d

4 4.75 m 2.06 m 0.71 d 0.85 d 7.49 d 4.91 m 2.68 d 3.10 d 6.89 d 6.64 d 9.12 s 2.70 s 4.60 dd 0.40 dt 1.54 dt 0.96 m 0.49 d 0.68 d 4.39 1.71 ddd m 2.30 m 1.71 m 1.71 m 4.88 s 7.35 d ---6.02 d 4.35 m 1.65 m 2.30 m 2.32 m 2.30 m 8.52 d

5 4.77 m 2.07 m 0.78 d 0.89 d 6.78 d 4.96 m 2.67 d 3.10 d 6.89 d 6.64 d 9.15 brs 2.71 s 4.64 dd 0.29 dt 1.62 dt 0.99 m 0.48 d 0.69 d 4.43 1.72 ddd m 2.35 m 1.59 m 2.03 m 4.40 s 7.20 d 3.01 s ---4.35 m 1.65 m 2.30 m 2.30 m 2.30 m 8.57 d

6 4.73 m 2.05 m 0.72 d 0.85 d 7.51 d 4.89 m 2.64 d 3.10 d 6.89 d 6.64 d 9.21 s 2.70 s 4.58 dd 0.40 dt 1.54 dt 0.96 m 0.48 d 0.69 d 4.38 1.71 ddd m 2.48 m 1.70 m 1.70 m 4.88 s 7.35 d ---6.02 d 4.34 m 1.61 m 2.22 m 2.21 m 2.18 m 8.49 d

7 4.77 m 2.07 m 0.78 d 0.89 d 6.75 d 4.96 m 2.67 d 3.10 d 6.89 d 6.62 d 9.15 brs 2.71 s 4.64 m 0.29 dt 1.62 m 0.99 m 0.48 d 0.69 d 4.43 m 1.72 m 2.35 m 1.59 m 2.03 m 4.40 s 7.20 d 3.01 s ---4.35 m 1.65 m 2.30 m 2.30 m 2.30 m 8.53 d

8 4.77 m 2.07 m 0.78 d 0.89 d 6.75 d 4.96 m 2.67 d 3.10 d 6.89 d 6.62 d 9.15 brs 2.71 s 4.64 m 0.29 dt 1.62 m 0.99 m 0.48 d 0.69 d 4.43 m 1.72 m 2.35 m 1.59 m 2.03 m 4.40 s 7.20 d 3.01 s ---4.35 m 1.65 m 2.30 m 2.30 m 2.30 m 8.53 d

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Thr

Tyr

Hpla

OMe NH2(a) NH2(b) 2 3 4 NH 2 3a 3b 5,5' 6,6' OH NH 2 3a 3b 5,5' 6,6' 2-OH 7-OH

---6.74 s 7.19 s 4.67 d 5.49 q 1.20 d 8.25 d 4.71 m 2.82 m 2.96 m 6.92 d 6.61 d 9.07 s 7.69 d 3.94 ddd 2.44 m 9.0,5.5,3.3 2.79 m 6.97 d 6.60 d 5.36 d 9.11 s

---7.21 d 6.75 s 4.70 d 5.53 q 1.21 d 8.29 d 4.72 m 2.81 m 2.96 m 6.93 d 6.62 d 9.14 s 7.71 d 3.96 2.44 ddd m 2.81 m 6.98 d 6.60 d 5.36 d 9.16 brs

3.56 s ------4.67 d 5.50 q 1.19 d 8.31 d 4.73 m 2.81 m 2.95 m 6.92 d 6.62 d 9.12 brs 7.69 d 3.94 2.45 ddd m 2.80 m 6.96 d 6.60 d 5.38 d 9.12 brs

3.56 s ------4.68 d 5.52 q 1.19 d 8.35 d 4.74 m 2.77 m 2.95 m 6.93 d 6.62 d 9.15 brs 7.69 d 3.96 2.45 ddd m 2.80 d 6.95 d 6.60 d 5.36 d 9.15 brs

---------4.67 d 5.49 q 1.19 d 8.28 d 4.70 m 2.77 m 2.95 m 6.92 d 6.62 d 9.08 s 7.67 d 3.95 2.43 ddd m 2.77 d 6.95 d 6.60 d 5.37 d 9.11 s

---------4.68 d 5.52 q 1.19 d 8.34 d 4.74 m 2.77 m 2.95 m 6.89 d 6.64 d 9.15 brs 7.67 d 3.96 2.45 ddd m 2.80 d 6.92 d 6.60 d 5.35 d 9.15 brs

---------4.68 d 5.52 q 1.19 d 8.34 d 4.74 m 2.77 m 2.95 m 6.89 d 6.64 d 9.15 brs 7.67 d 3.96 2.45 ddd m 2.80 d 6.92 d 6.60 d 5.35 d 9.15 brs

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S3. Table S2. 13C NMR Data of 1 and its isolation artifacts 3-8 in DMSO-d6 Position Val

N-MeTyr

Leu

Ahp/Amp

Gln/Glu/ OMe Glu

1 2 3 4 5 1 2 3 4 5,5' 6,6' 7 N Me 1 2 3 4 5 6 2 3 4 5 6 O Me 1 2

1 δC [ppm] s 172.5 56.2 d 30.9 d 17.8 q 19.5 q 169.3 s 61.1 d 33.1 t 127.5 s 130.5 d 115.5 d 156.2 s 30.6 q 171.0 s 47.9 d 38.7 t 23.8 d 22.3 q 24.1 q 169.7 s 49.2 d 22.0 t 30.0 t 73.6 d ---170.2 s 52.3 d

3 δC [ppm] s 172.3 56.0 d 31.3 d 17.8 q 19.4 q 169.3 s 61.3 d 33.0 t 127.4 s 130.2 d 115.0 d 156.2 s 30.5 q 170.9 s 47.4 d 38.0 t 23.8 d 22.1 q 24.0 q 169.1 s 49.1 d 22.0 t 23.8 t 82.6 d 55.5 q 170.3 s 52.1 d






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Thr

Tyr

Hpla

3 4 CO O Me 1 2 3 4 1 2 3 4 5,5' 6,6' 7 1 2 3 4 5,5' 6,6' 7

27.0 t 31.9 t 173.9 s ---169.3 s 55.0 d 72.1 d 17.5 q 171.9 s 53.3 d 37.0 t 127.4 s 130.4 d 115.0 d 155.7 s 173.3 s 72.5 d 39.8 t 128.9 s 130.2 d 114.8 d 155.9 s


26.1 t 30.0 t 173.0 s 51.6 q 169.3 s 55.0 d 72.1 d 17.4 q 171.8 s 53.2 d 37.1 t 127.4 s 130.2 d 115.5 d 156.2 s 173.2 s 72.5 d 39.1 t 128.8 s 130.4 d 114.9 d 155.7 s

26.1 t 30.0 t 173.0 s 51.6 q 169.4 s 55.0 d 72.1 d 17.4 q 171.9 s 53.2 d 37.1 t 127.4 s 130.3 d 115.7 d 156.3 s 173.3 s 72.6 d 39.2 t 128.9 s 130.5 d 114.9 d 155.8 s



26.3 t 30.0 t 173.0 s ---169.4 s 55.0 d 72.1 d 17.4 q 171.9 53.2 d 37.1 t 127.4 s 130.2 d 115.6 d 156.3 s 173.3 s 72.5 d 39.2 t 128.9 s 130.4 d 114.9 d 155.7 s

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RH-100.2.1.ﬁd RH-100.2/ DMSO 29.10.2014 9.20 9.11 9.07 8.51 8.49 8.26 8.24 7.70 7.69 7.55 7.53 7.35 7.33 7.20 7.19 7.17 7.07 6.99 6.98 6.97 6.97 6.93 6.93 6.92 6.90 6.88 6.75 6.74 6.62 6.61 6.60 6.59 6.51 6.03 6.02 5.51 5.49 5.48 5.37 5.36 4.91 4.89 4.74 4.73 4.72 4.71 4.70 4.68 4.66 4.61 4.60 4.59 4.58 4.40 4.38 4.36 4.35 4.32 4.31 4.30 4.29 4.28 4.27 4.08 3.94 3.32 HDO 3.16 3.11 3.08 2.98 2.97 2.95 2.82 2.79 2.77 2.76 2.70 2.68 2.65 2.52 2.50 DMSO 2.49 DMSO 2.49 DMSO 2.49 DMSO 2.48 DMSO 2.47 2.45 2.44 2.42 2.19 2.17 2.14 2.11 2.09 2.08 2.06 2.05 2.04 2.02 2.00 1.99 1.90 1.71 1.68 1.67 1.65 1.56 1.54 1.52 1.22 1.21 1.20 0.97 0.86 0.84 0.74 0.72 0.69 0.68 0.50 0.48 0.44 0.42 0.40

S4. 1H NMR Spectrum of Micropeptin TR1058 (1) in DMSO-d6

10.5 10.0 9.5 9.0 8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 f1 (ppm) 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0 -0.5

550

500

450

400

350

300

250

200

150

100

50

0

-1.0 -50

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S5. 13C NMR Spectrum of Micropeptin TR1058 (1) in DMSO-d6

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S6. HSQC Spectrum Micropeptin TR1058 (1) in DMSO-d6 (CH, CH3, CH2)

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S7. HMBC Spectrum of Micropeptin TR1058 (1) in DMSO-d6

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S8. COSY Spectrum of Micropeptin TR1058 (1) in DMSO-d6

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S9. TOCSY Spectrum of Micropeptin TR1058 (1) in DMSO-d6

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S10. ROESY Spectrum of Micropeptin TR1058 (1) in DMSO-d6 (NOE correlation)

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S11. HR ESI MS data of Micropeptin TR1058 (1)

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S12. 1H NMR Spectrum of Aeruginosin TR642 (2) in DMSO-d6

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S13. 13C NMR Spectrum of Aeruginosin TR642 (2) in DMSO-d6

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S14. HSQC Spectrum of Aeruginosin TR642 (2) in DMSO-d6 (CH, CH3, CH2)

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S15. HMBC Spectrum of Aeruginosin TR642 (2) in DMSO-d6

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S16. COSY Spectrum of Aeruginosin TR642 (2) in DMSO-d6

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S17. TOCSY Spectrum of Aeruginosin TR642 (2) in DMSO-d6

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S18. ROESY Spectrum of Aeruginosin TR642 (2) in DMSO-d6 (NOE correlation)

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S19. HR ESI MS data of Aeruginosin TR642 (2)

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