Thiolactone modulators of quorum sensing revealed through ... · Thiolactone modulators of quorum sensing revealed through library design and screening Christine E. McInnis and Helen

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Thiolactone modulators of quorum sensing revealed through

library design and screening

Christine E. McInnis and Helen E. Blackwell∗

Department of Chemistry, University of Wisconsin–Madison, 1101 University Avenue,

Madison, WI 53706-1322

E-mail: [email protected]

Supplementary Material. General instrumentation and analytical methods................................................................... S-2 GC purity data for thiolactone analogs .................................................................................. S-3 HPLC purity data for thiolactone analogs ............................................................................. S-6 Compound characterization data............................................................................................ S-8 Biological screening protocols and supplementary assay data .............................................. S-11 References.............................................................................................................................. S-15

∗ Corresponding author. Phone: 608/262-1503; Fax: 608/265-4534

McInnis and Blackwell Supplementary Material

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General instrumentation and analytical methods. All chemical reagents were purchased from commercial sources (Alfa-Aesar, Aldrich, and Acros) and used without further purification. Solvents were purchased from commercial sources (Aldrich and J.T. Baker) and used as obtained, with the exception of dichloromethane (CH2Cl2), which was distilled over calcium hydride immediately prior to use. 1H and 13C NMR spectra were recorded on a Bruker AC-300 spectrometer in deuterated chloroform at 300 MHz. Chemical shifts are reported in parts per million (ppm). Tetramethyl silane (TMS) was used as an internal reference (0.0 ppm) for 1H NMR spectra. The chloroform carbons were used as an internal reference for 13C NMR spectra. Couplings are reported in hertz. Electrospray ionization (ESI) MS data were obtained using a Shimadzu LCMS-2010 system equipped with two pumps (LC-10ADvp), a controller (SCL-10Avp), an autoinjector (SIL-10Avp), a UV diode array detector (SPD-M10Avp), and a single quadrupole analyzer. Electron impact (EI) MS data were obtained on a Shimadzu single quadrupole GCMS-QP2010S. Optical rotations (αD

25) were measured on a Perkin-Elmer 241 digital polarimeter at 25 °C. Infrared (IR) spectra were measured on a Bruker Equinox 55 ATR-FT-IR using a germanium crystal. Gas chromatography (GC) data were obtained using a Shimadzu GC-2010 system. A Shimadzu SHRXI-5MS capillary column (dimensions: 30 m x 0.25 mm x 0.25 μm) was used for all GC work. The standard GC method was as follows: injection temperature 275 °C; initial oven temperature 100 °C; hold 5 min; ramp at 10 °C/min to 300 °C; hold 5 min, for a total run time of 30 min. GC could not be used to characterize selected thiolactones (e.g., 3-oxo derivatives) due to decomposition at high temperature; reversed-phase high performance liquid chromatography (RP-HPLC) was therefore used to characterize these compounds. RP-HPLC data were obtained using a Shimadzu HPLC instrument equipped with a single pump (LC-10Atvp), a solvent mixer (FCV-10Alvp), a controller (SCL-10Avp), an autoinjector (SIL-10AF), and a UV diode array detector (SPD-M10Avp). A Shimadzu Premier C-18 RP column (dimensions: 25 cm x 4.6 mm) was used for all HPLC work. The standard HPLC method was to ramp from 70–95% acetonitrile in water over 20 min.


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GC purity data for thiolactone analogs. The major compound peak is marked with an asterisk.

Compound Purity Structure HPLC chromatogram

DMSO blank --

19 93%

20 97%

*

*


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21 92%

22 ~100%

23 91%

*

*

*


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27 98%

29 ~100%

31 97%

*

*

*


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33 99%

HPLC purity data for thiolactone analogs.

Compound Purity Structure HPLC chromatogram

Methanol blank --

12 98%

15 97%

*


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18 97%

25 98%

29 99%

35 ~100%


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Compound characterization data. 1H and 13C NMR, ESI MS, IR, and [α]D data are reported below for all of the L-thiolactone analogs. The D,L-thiolactone analogs gave analogous NMR, MS, and IR data (not shown). 12: 1H NMR: (300 MHz, CDCl3) δ 7.47 (NH), 4.59 (CH-lac, ddd, J = 13.0, 7.2, 6.7 Hz, 1H), 3.45 (CH2, s, 2H), 3.36 (CH-lac, ddd (apparent td), J = 11.4, 5.2 Hz, 1H), 3.26 (CH-lac, ddd, J = 11.4, 10.8, 1.4 Hz, 1H), 2.88 (CH-lac, ddd, J = 12.4, 5.1, 1.4 Hz, 1H), 2.52 (CH2, t, J = 7.2 Hz, 2H), 2.02 (CH-lac, ddd (apparent dt), J = 12.5, 7.1 Hz, 1H), 1.71 (CH2, m (apparent broad singlet), 2H), 1.63 (CH2, AB q, J = 7.4 Hz, 2H), 0.93 (CH3, t, J = 7.5 Hz, 3H); 13C NMR: (300 MHz, CDCl3) δ 206.6, 204.9, 166.5, 59.5, 48.6, 45.9, 31.7, 27.7, 17.0, 13.7; ESI: expected m/z = 229.3, observed [M+Na+] = 252.2; IR (cm-1): 3263, 2965, 1698, 1646, 1555, 1057, 907; [α]D (CHCl3, c = 4.4 mg/mL): +57.1° 15: 1H NMR: (300 MHz, CDCl3) δ 7.55 (NH), 4.64 (CH-lac, ddd, J = 12.8, 7.1, 6.8 Hz, 1H), 3.51 (CH2, s, 2H), 3.41 (CH-lac, ddd (apparent td), J = 11.6, 5.3 Hz, 1H), 3.31 (CH-lac, ddd, J = 11.3, 10.9, 1.1 Hz, 1H), 2.91 (CH-lac, ddd, J = 12.4, 5.3, 1.1 Hz, 1H), 2.57 (CH2, t, J = 7.5 Hz, 2H), 2.07 (CH-lac, ddd (apparent dt), J = 12.4, 6.8 Hz, 1H), 1.65 (CH2, m, 2H), 1.31 (CH2, m (broad s), 12H), 0.93 (CH3, t, J = 5.6 Hz, 3H); 13C NMR: (300 MHz, CDCl3) δ 206.8, 204.6, 166.4, 59.5, 48.5, 44.2, 32.0, 31.7, 29.6, 29.5, 29.4, 29.2, 27.7, 23.6, 22.9, 14.3; ESI: expected m/z = 313.5, observed [M+Na+] = 336.3; IR (cm-1): 3262, 2924, 2854, 1699, 1647, 1557, 1350, 1055, 927; [α]D (CHCl3, c = 2.7 mg/mL): +43.9° 18: 1H NMR: (300 MHz, CDCl3) δ 7.45 (NH), 4.58 (CH-lac, ddd (apparent dt), J = 12.6, 6.8 Hz, 1H), 3.45 (CH2, s, 2H), 3.36 (CH-lac, ddd (apparent td), J = 11.6, 5.8 Hz, 1H), 3.26 (CH-lac, ddd, J = 11.0, 11.0, 1.1 Hz, 1H), 2.86 (CH-lac, ddd, J = 12.6, 5.1, 1.2 Hz, 1H), 2.52 (CH2, t, J = 7.3 Hz, 2H), 2.01 (CH-lac, ddd (apparent dt), J = 12.0, 7.1 Hz, 1H), 1.60 (CH2, m, 2H), 1.30 (CH2, m, 4H), 0.89 (CH3, t, J = 6.9 Hz, 3H); 13C NMR: (300 MHz, CDCl3) δ 206.6, 204.9, 166.5, 59.4, 48.7, 44.0, 31.6, 31.3, 27.7, 23.2, 22.6, 14.1; ESI: expected m/z =257.4, observed [M+Na+] = 280.2; IR (cm-1): 3271, 2931, 1699, 1645, 1559, 1417, 1343, 1056, 921; [α]D (CHCl3, c = 12.2 mg/mL): +51.2° 19: 1H NMR: (300 MHz, CDCl3) δ 5.97 (NH), 4.53 (CH-lac, ddd, J = 12.9, 7.3, 6.2 Hz, 1H), 3.37 (CH-lac, ddd (apparent td), J = 11.7, 5.1 Hz, 1H), 3.26 (CH-lac, ddd, J = 11.5, 11.0, 1.1 Hz, 1H), 2.96 (CH-lac, ddd, J = 12.4, 5.1, 1.3 Hz, 1H), 2.23 (CH2, t, J = 7.6 Hz, 2H), 1.92 (CH-lac, ddd (apparent dt), J = 12.5, 7.0 Hz, 1H), 1.68 (CH2, dq, J = 15.7, 7.3 Hz, 2H), 0.96 (CH3, t, J = 7.4 Hz, 3H); 13C NMR: (300 MHz, CDCl3) δ 205.9, 173.7, 59.7, 38.5, 32.3, 27.8, 19.2, 13.9; ESI: expected m/z = 187.1, observed [M+Na+] = 210.1; IR (cm-1): 3266, 1701, 1647, 1546, 1056, 1021, 921; [α]D (CHCl3, c = 6.8 mg/mL): +59.3° 20: 1H NMR: (300 MHz, CDCl3) δ 6.03 (NH), 4.53 (CH-lac, ddd, J = 12.5, 7.1, 5.4 Hz, 1H), 3.37 (CH-lac, ddd (apparent td), J = 11.6, 5.4 Hz, 1H), 3.25 (CH-lac, ddd, J = 11.1, 7.2, 0.9 Hz, 1H), 2.95 (CH-lac, ddd, J = 12.4, 5.0, 1.0 Hz, 1H), 2.24 (CH2, t, J = 8.1 Hz, 2H), 1.92 (CH-lac, ddd (apparent dt), J = 12.4, 6.9 Hz, 1H), 1.65 (CH2, m, 2H), 1.32 (CH2, m, 4H), 0.90 (CH3, t, J =


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7.0 Hz, 3H); 13C NMR: (300 MHz, CDCl3) δ 206.0, 174.0, 59.7, 36.6, 32.3, 31.6, 27.8, 25.4, 22.6, 14.1; ESI: expected m/z = 215.1, observed [M+Na+] = 238.2; IR (cm-1): 3284, 2933, 1694, 1641, 1533, 913; [α]D (CHCl3, c = 11.6 mg/mL): +32.6° 21: 1H NMR: (300 MHz, CDCl3) δ 5.83 (NH), 4.49 (CH-lac, ddd, J = 12.7, 6.6, 6.1 Hz, 1H), 3.36 (CH-lac, ddd (apparent td), J = 11.5, 5.1 Hz, 1H), 3.25 (CH-lac, ddd, J = 11.3, 7.1, 0.8 Hz, 1H), 2.99 (CH-lac, ddd, J = 12.3, 5.1, 1.2 Hz, 1H), 2.24 (CH2, t, J = 7.3 Hz, 2H), 1.89 (CH-lac, ddd (apparent dt), J = 12.3, 7.1 Hz, 1H), 1.64 (CH2, m, 2H), 1.30 (CH2, m, 6H), 0.88 (CH3, t, J = 6.8 Hz, 3H); 13C NMR: (300 MHz, CDCl3) δ 205.9, 173.9, 59.7, 36.6, 32.4, 31.7, 29.1, 27.8, 25.7, 22.7, 14.2; ESI: expected m/z = 230.1, observed 230.3; IR (cm-1): 3271, 2929, 2856, 1690, 1643, 1548, 1535, 1022, 917; [α]D (CHCl3, c = 7.5 mg/mL): +45.2° 22: 1H NMR: (300 MHz, CDCl3) δ 5.83 (NH), 4.49 (CH-lac, ddd, J = 13.0, 7.0, 6.0 Hz, 1H), 3.36 (CH-lac, ddd (apparent td), J = 11.5, 5.1 Hz, 1H), 3.25 (CH-lac, ddd, J = 11.6, 7.1, 1.1 Hz, 1H), 2.99 (CH-lac, ddd, J = 12.3, 5.0, 1.1 Hz, 1H), 2.24 (CH2, t, J = 7.1 Hz, 2H), 1.89 (CH-lac, ddd (apparent dt), J = 11.3, 7.1 Hz, 1H), 1.64 (CH2, m, 2H), 1.29 (CH2, m, 8H), 0.88 (CH3, t, J = 6.9 Hz, 2H); 13C NMR: (300 MHz, CDCl3) δ 205.9, 173.9, 59.7, 36.6, 32.4, 31.9, 29.4, 29.2, 27.8, 25.7, 22.8, 14.3; ESI: expected m/z = 243.1, observed [M+Na+] = 266.2; IR (cm-1): 3272, 2930, 2856, 1688, 1643, 1547, 1209, 919; [α]D (CHCl3, c = 5.3 mg/mL): +41.4° 23: 1H NMR: (300 MHz, CDCl3) δ 5.84 (NH), 4.49 (CH-lac, ddd, J = 12.9, 6.4, 5.8 Hz, 1H), 3.36 (CH-lac, ddd (apparent td), J = 11.6, 5.2 Hz, 1H), 3.25 (CH-lac, ddd, J = 12.0, 11.2, 1.0 Hz, 1H), 2.99 (CH-lac, ddd, J = 12.6, 5.2, 1.2 Hz, 1H), 2.24 (CH2, t, J = 7.4 Hz, 2H), 1.89 (CH-lac, ddd (apparent dt), J = 12.6, 7.0 Hz, 1H), 1.64 (CH2, m, 2H), 1.27 (CH2, m, 16H), 0.88 (CH3, t, J = 7.0 Hz, 3H); 13C NMR: (300 MHz, CDCl3) δ 206.0, 174.0, 59.7, 36.6, 34.1, 32.3, 32.1, 29.8, 29.7, 29.5, 29.4, 27.8, 25.7, 25.0, 22.9, 14.3; ESI: expected m/z = 299.2, observed [M+Na+] = 322.4; IR (cm-1):3281, 2919, 2851, 1697, 1646, 1549, 1473, 911; [α]D (CHCl3, c = 9.5 mg/mL): +26.1° 25: 1H NMR: (300 MHz, CDCl3) δ 7.32 (Ar-H, m, 5H), 5.91 (NH), 4.51 (CH-lac, ddd (apparent td), J = 13.0, 6.4 Hz, 1H), 3.63 (CH2, s, 2H), 3.34 (CH-lac, ddd (apparent td), J = 11.7, 5.0 Hz, 1H), 3.22 (CH-lac, ddd, J = 11.1, 7.2, 0.8 Hz, 1H), 2.90 (CH-lac, ddd, J = 12.7, 5.0, 1.1 Hz, 1H), 1.87 (CH-lac, ddd (apparent dt), J = 13.0, 7.2 Hz, 1H); 13C NMR: (300 MHz, CDCl3) δ 205.3, 171.7, 134.4, 129.6, 129.3, 127.7, 59.7, 43.7, 32.0, 27.7; ESI: expected m/z = 235.1, observed [M+Na+] = 258.2; IR (cm-1): 3266, 3068, 1695, 1651, 1544, 700; [α]D (CHCl3, c = 7.7 mg/mL): +20.6° 27: 1H NMR: (300 MHz, CDCl3 ) δ 8.16 (Ar-H, m, 2H), 7.65 (Ar-H, d, J = 7.4 Hz, 1H), 7.54 (Ar-H, t, J = 8.4 Hz, 1H), 5.98 (NH), 4.49 (CH-lac, ddd, J = 13.5, 7.9, 6.5 Hz, 1H), 3.70 (CH2, s, 2H), 3.36 (CH-lac, ddd (apparent td), J = 11.6, 5.0 Hz, 1H), 3.26 (CH-lac, ddd, J = 11.9, 11.6, 1.0 Hz, 1H), 2.97 (CH-lac, ddd, J = 12.5, 5.4, 1.5 Hz, 1H), 1.91 (CH-lac, ddd (apparent dt), J = 12.3, 6.9 Hz, 1H); 13C NMR: (300 MHz, CDCl3) δ 205.5, 170.2, 148.6, 136.4, 135.8, 129.9, 124.5, 122.6, 59.9, 42.6, 32.0, 27.8; ESI: expected m/z = 280.3, observed [2M+Na+] = 583.3; IR


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(cm-1): 3270, 3079, 2928, 2360, 1694, 1647, 1558, 1519, 1348, 907, 807, 728, 672; [α]D (CHCl3, c = 9.3 mg/mL): +41.7° 29: 1H NMR: (300 MHz, CDCl3) δ 7.19 (Ar-H, d, J = 9.3 Hz, 2H), 6.89 (Ar-H, d, J = 8.7 Hz, 2H), 5.86 (NH), 4.50 (CH-lac, ddd, J = 13.3, 7.0, 6.4 Hz, 1H), 3.80 (OCH3, s, 3H), 3.56 (CH2, s, 2H), 3.33 (CH-lac, ddd (apparent td), J = 11.7, 5.3 Hz, 1H), 3.21 (CH-lac, ddd, J = 11.0, 7.3, 1.1 Hz, 1H), 2.88 (CH-lac, ddd, J = 12.6, 5.1, 1.1 Hz, 1H), 1.85 (CH-lac, ddd (apparent dt), J = 12.1, 7.3 Hz, 1H); 13C NMR: (300 MHz, CDCl3) δ 205.3, 172.1, 159.2, 130.7, 126.3, 114.7, 59.7, 55.5, 42.8, 32.0, 27.7; ESI: expected m/z = 265.3, observed [M+Na+] = 288.2; IR (cm-1): 3248, 3061, 1688, 1550, 1517, 1244, 1024, 821; [α]D (CHCl3, c = 1.2 mg/mL): +8.9° 31: 1H NMR: (300 MHz, CDCl3) δ 7.48 (Ar-H, d, J = 8.4 Hz, 2H), 7.16 (Ar-H, d, J = 8.3 Hz, 2H), 5.810 (NH), 4.48 (CH-lac, ddd, J = 12.7, 6.7, 6.4 Hz, 1H), 3.341 (CH-lac, ddd (apparent dt), J = 11.7, 5.1 Hz, 1H), 3.56 (CH2, s, 2H), 3.23 (CH-lac, ddd, J = 11.4, 7.1, 0.9 Hz, 1H), 2.92 (CH-lac, ddd, J = 12.4, 5.0, 1.2 Hz, 1H), 1.86 (CH-lac, ddd (apparent td), J = 12.6, 7.3 Hz, 1H); 13C NMR: (300 MHz, CDCl3) δ 205.4, 171.0, 133.4, 132.3, 131.2, 121.9, 59.7, 42.9, 31.9, 27.4; ESI: expected m/z = 314.0, observed [M+Na+] = 337.1; IR (cm-1): 3274, 2925, 2360, 1700, 1654, 1538, 1487, 1012, 909; [α]D (CHCl3, c = 10.6 mg/mL): +27.1° 33: 1H NMR: (300 MHz, CDCl3) δ 7.59 (Ar-H, d, J = 7.8 Hz, 4H), 7.44 (Ar-H, m, 2H), 7.35 (Ar-H, m, 3H), 5.95 (NH), 4.52 (CH-lac, ddd, J = 12.8, 6.4, 6.4 Hz, 1H), 3.66 (CH2, s, 2H), 3.34 (CH-lac, ddd, J = 11.3, 11.3, 4.9 Hz, 1H), 3.22 (CH-lac, ddd, J = 11.5, 6.9, 0.7 Hz, 1H), 2.91 (CH-lac, ddd, J = 12.3, 5.0, 1.0 Hz, 1H), 1.88 (CH-lac, ddd, J = 12.2, 12.2, 7.1 Hz, 1H); 13C NMR: (300 MHz, CDCl3) δ 205.4, 171.6, 140.7, 133.4, 130.1, 129.0, 128.0, 127.6, 127.3, 59.8, 43.3, 32.0, 27.7; ESI: expected m/z = 311.1, observed [M+Na+] = 334.2; IR (cm-1): 3267, 3059, 2929, 1696, 1648, 1543, 1489, 1262, 914, 752, 690; [α]D (CHCl3, c = 6.5 mg/mL): +25.1° 35: 1H NMR: (300.1 MHz, CDCl3) δ 8.04 (NH), 7.60 (Ar-H, d, J = 7.4 Hz, 1H), 7.34 (Ar-H, d, J = 8.4 Hz, 1H), 7.15 (Ar-H, m, 2H), 6.99 (Ar-H, d, J = 2.6 Hz, 1H), 5.87 (NH), 4.48 (CH-lac, ddd (apparent dt), J = 12.7, 6.9 Hz, 1H), 3.28 (CH-lac, ddd (apparent td), J = 11.6, 5.3 Hz, 1H), 3.16 (CH-lac, ddd, J = 10.0, 6.9, 2.1 Hz, 1H), 3.11 (CH2, t, J = 8.4 Hz, 2H), 2.79 (CH-lac, ddd, J = 12.6, 5.3, 1.5 Hz, 1H), 2.62 (CH2, td, J = 7.8, 1.9 Hz, 2H), 1.74 (CH-lac, ddd (apparent dt), J = 12.9, 6.9 Hz, 1H); 13C NMR: (75.4 MHz, CDCl3) δ 205.8, 173.6, 136.6, 127.3, 122.0, 121.6, 119.6, 118.9, 114.9, 111.5, 59.6, 37.3, 32.0, 27.7, 21.3; ESI: expected m/z = 288.4, observed [M+Na+] = 311.2; IR (cm-1): 3386, 2934, 1694, 1656, 1527, 1344, 1059, 1020, 918, 745; [α]D (CHCl3, c = 7.9 mg/mL): +38.5°


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Biological screening protocols and supplementary assay data. Compound handling and reagents. Stock solutions of synthetic compounds (10 mM) were prepared in DMSO and stored at room temperature (rt) in sealed vials. Solvent resistant polypropylene (Corning Costar cat. no. 3790), clear polystyrene (Corning Costar cat. no. 3997), black polystyrene clear bottom (Corning Costar cat. no. 3603), or white polystyrene clear bottom (Corning Costar cat. no. 3610) 96-well microtiter plates were used as appropriate. All biological reagents were purchased from Fisher and used according to enclosed instructions. Buffers, media, and solutions were prepared as previously described[1] for E. coli DH5α (pJN105L + pSC11),[2] V. fischeri ESI 114 (Δ-LuxI),[3] and A. tumefaciens WCF (pCF372).[4] Medium for assays using P. aeruginosa PA01 MW1 (pUM15)[5] was prepared as follows: 20 g/L Luria-Bertani (LB) broth (Sigma) and 10.4 g/L 3-(N-morpholino)propanesulfonic acid (MOPS, Acros) were dissolved in distilled water, and the pH was adjusted to 7 prior to autoclaving. Immediately before using the medium, carbenicillin (300 μg/mL, Sigma) was added. Instrumentation. Absorbance and fluorescence measurements were obtained using a PerkinElmer Wallac 2100 EnVision multilabel plate reader running Wallac Manager v1.03 software. A 600 nm filter was used for reading bacterial cell density. Filters of 420 nm and 550 nm were used for Miller-type absorbance assays. Filters of 485 nm (excitation) and 535 nm (emission) were used for evaluating the production of yellow fluorescent protein (YFP) in fluorescence assays. Reporter gene assays. Reporter gene assays for E. coli DH5α (pJN105L + pSC11), V. fischeri ESI 114 (Δ-LuxI), and A. tumefaciens WCF (pCF372) were conducted as previously described.[1] The assay using P. aeruginosa PA01 MW1 (pUM15) was modified from a literature procedure.[5] In brief, cultures were prepared by adding one PA01 MW1 colony to 5 mL of prepared medium and growing the culture overnight at 30 °C. A subculture was prepared the following morning (~16 h later) by making a 1:10 dilution of the overnight culture. This subculture was incubated with shaking at 37 °C until it reached an optical density of 0.25–0.30, after which it was plated into microtiter plate wells in 200 μL aliquots. Thiolactones were added directly to the wells to achieve 10 μM concentrations. For antagonism assays, OdDHL was also added to the wells to give a final concentration of 1 μM; agonists assays contained only the thiolactone. The outer wells of the compound plate were filled with subculture minus compound to circumvent problems due to evaporation. After plating was complete, plates were incubated with shaking at 37 °C for 8 h. The optical density and fluorescence of the wells were read thereafter. Dose response curves. Dose response analyses for active thiolactones were performed in triplicate of triplicate using the reporter strains above over a range of concentrations. Error bars are s. d. of triplicate samples. EC50 and IC50 values were calculated using Graph Pad Prism software (version 4, for Macintosh). The dose responses curves are shown in Figures S-1–S-4 below.


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Figure S-1. Dose response curves for thiolactones 12, 15, 20–23, 25, 27, 29, 31, 33, and 35 in the E. coli strain (LasR reporter).


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Figure S-2. Dose response curves for thiolactones 12, 15, and 23 in the P. aeruginosa strain (LasR reporter).

Figure S-3. Dose response curves for thiolactones 15, 18, 20–22, 27, 31, and 33 in the V. fischeri strain (LuxR reporter).


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Figure S-4. Dose response curves for thiolactones 12, 15, 18, 21, and 22 in the A. tumefaciens strain (TraR reporter).


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References. [1] G. D. Geske, J. C. O'Neill, D. M. Miller, M. E. Mattmann, H. E. Blackwell, J. Am. Chem.

Soc. 2007, 129, 13613.

[2] J. H. Lee, Y. Lequette, E. P. Greenberg, Mol. Microbiol. 2006, 59, 602.

[3] C. Lupp, M. Urbanowski, E. P. Greenberg, E. G. Ruby, Mol. Microbiol. 2003, 50, 319.

[4] J. Zhu, J. W. Beaber, M. I. More, C. Fuqua, A. Eberhard, S. C. Winans, J. Bacteriol. 1998, 180, 5398.

[5] U. Muh, M. Schuster, R. Heim, A. Singh, E. R. Olson, E. P. Greenberg, Antimicrob. Agents Chemother. 2006, 50, 3674.

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Thiolactone modulators of quorum sensing revealed through ... · Thiolactone modulators of quorum sensing revealed through library design and screening Christine E. McInnis and Helen