Journal of the American Chemical Society
Article
anaerobic and aerobic cultures, respectively. For 96-well plates, each
well contained 90 μL of an overnight culture containing L. lactis HP
(approximately 1 × 108 CFU mL−1) and a 10× stock of peptide (10
μL) of the desired concentration. The plates were incubated at 30 °C
overnight with no agitation. For 48-well plates, each well contained
180 μL of an overnight culture containing B. subtilis 168
(approximately 1 × 108 CFU mL−1) and a 10× stock of peptide (20
μL) of the desired concentration. The plates were incubated at 30 °C
overnight with moderate shaking. In addition, each 96- and 48-well
plate contained several blank (growth media with no bacteria) and
control (SDW in place of peptide) wells. The optical density at 630
nm (OD630) was recorded at hourly intervals from 0 to 6 h and a final
recording at 22−24 h. For experiments with Cy5-Halβ, the optical
density was read at 570 nm (OD570) to minimize the undesired
excitation of Cy5. The MIC was determined as the lowest
concentration at which no cell growth was observed after 22−24 h.
(10) Knerr, P. J.; van der Donk, W. A. J. Am. Chem. Soc. 2013, 135,
7094.
(11) Cotter, P. D.; Hill, C.; Ross, R. P. Curr. Protein Pept. Sci. 2005, 6,
61.
(12) van Heel, A. J.; Montalban-Lopez, M.; Kuipers, O. P. Expert
Opin. Drug Metab. Toxicol. 2011, 7, 675.
(13) Kuipers, O. P.; Bierbaum, G.; Ottenwalder, B.; Dodd, H. M.;
̈
Horn, N.; Metzger, J.; Kupke, T.; Gnau, V.; Bongers, R.; van den
Bogaard, P.; Kosters, H.; Rollema, H. S.; de Vos, W. M.; Siezen, R. J.;
Jung, G.; Gotz, F.; Sahl, H. G.; Gasson, M. J. Antonie van Leeuwenhoek
̈
1996, 69, 161.
(14) Ross, A. C.; Vederas, J. C. J. Antibiot. 2011, 64, 27.
(15) Field, D.; Hill, C.; Cotter, P. D.; Ross, R. P. Mol. Microbiol.
2011, 78, 1077.
(16) Cortes, J.; Appleyard, A. N.; Dawson, M. J. Methods Enzymol.
́
2009, 458, 559.
(17) Szekat, C.; Jack, R. W.; Skutlarek, D.; Farber, H.; Bierbaum, G.
Appl. Environ. Microbiol. 2003, 69, 3777.
ASSOCIATED CONTENT
■
(18) Asaduzzaman, S. M.; Nagao, J.; Aso, Y.; Nakayama, J.;
Sonomoto, K. Appl. Environ. Microbiol. 2006, 72, 6012.
(19) Cooper, L. E.; McClerren, A. L.; Chary, A.; van der Donk, W. A.
Chem. Biol. 2008, 15, 1035.
(20) Islam, M. R.; Shioya, K.; Nagao, J.; Nishie, M.; Jikuya, H.;
Zendo, T.; Nakayama, J.; Sonomoto, K. Mol. Microbiol. 2009, 72, 1438.
(21) Levengood, M. R.; Knerr, P. J.; Oman, T. J.; van der Donk, W.
A. J. Am. Chem. Soc. 2009, 131, 12024.
(22) Deegan, L. H.; Suda, S.; Lawton, E. M.; Draper, L. A.;
Hugenholtz, F.; Peschel, A.; Hill, C.; Cotter, P. D.; Ross, R. P. Microb.
Biotechnol. 2010, 3, 222.
(23) Oman, T. J.; Lupoli, T. J.; Wang, T. S.; Kahne, D.; Walker, S.;
van der Donk, W. A. J. Am. Chem. Soc. 2011, 133, 17544.
(24) Knerr, P. J.; Oman, T. J.; Garcia De Gonzalo, C. V.; Lupoli, T.
J.; Walker, S.; van der Donk, W. A. ACS Chem. Biol. 2012, 7, 1791.
(25) Suda, S.; Hill, C.; Cotter, P. D.; Ross, R. P. Bioeng. Bugs 2010, 1,
345.
S
* Supporting Information
Experimental procedures for plasmid construction, purification
and characterization of LanA analogues, as well as supporting
figures. This material is available free of charge via the Internet
AUTHOR INFORMATION
■
Corresponding Author
(217) 244-8533.
Notes
The authors declare no competing financial interest.
ACKNOWLEDGMENTS
■
This work was supported by the National Institutes of Health
(RO1 GM058822). We thank Dr. Rebecca Splain for providing
Figure 7.
(26) Stephanopoulos, N.; Francis, M. B. Nat. Chem. Biol. 2011, 7,
876.
(27) Chen, I.; Howarth, M.; Lin, W.; Ting, A. Y. Nat. Methods 2005,
2, 99.
(28) Carrico, I. S.; Carlson, B. L.; Bertozzi, C. R. Nat. Chem. Biol.
REFERENCES
■
2007, 3, 321.
(1) Arnison, P. G. B., M. J.; Bierbaum, G.; Bowers, A. A.; Bulaj, G.;
Camarero, J. A.; Campopiano, D. J.; Clardy, J.; Cotter, P. D.; Craik, D.
J.; Dawson, M.; Dittmann, E.; Donadio, S.; Dorrestein, P. C.; Entian,
(29) Griffin, B. A.; Adams, S. R.; Tsien, R. Y. Science 1998, 281, 269.
(30) Yin, J.; Liu, F.; Li, X.; Walsh, C. T. J. Am. Chem. Soc. 2004, 126,
7754.
(31) Gautier, A.; Juillerat, A.; Heinis, C.; Correa, I. R., Jr.;
Kindermann, M.; Beaufils, F.; Johnsson, K. Chem. Biol. 2008, 15, 128.
(32) Wagner, A. M.; Fegley, M. W.; Warner, J. B.; Grindley, C. L.;
Marotta, N. P.; Petersson, E. J. J. Am. Chem. Soc. 2011, 133, 15139.
(33) Antos, J. M.; Chew, G. L.; Guimaraes, C. P.; Yoder, N. C.;
Grotenbreg, G. M.; Popp, M. W.; Ploegh, H. L. J. Am. Chem. Soc. 2009,
131, 10800.
K.-D.; Fischbach, M. A.; Garavelli, J. S.; Goransson, U.; Gruber, C. W.;
̈
Haft, D. H.; Hemscheidt, T. K.; Hertweck, C.; Hill, C.; Horswill, A. R.;
Jaspars, M.; Kelly, W. L.; Klinman, J. P.; Kuipers, O. P.; Link, A. J.; Liu,
W.; Marahiel, M. A.; Mitchell, D. A.; Moll, G. N.; Moore, B. S.; Nair, S.
K.; Nes, I. F.; Norris, G. E.; Olivera, B. M.; Onaka, H.; Patchett, M. L.;
Reaney, M. J. T.; Rebuffat, S.; Ross, R. P.; Sahl, H.-G.; Saris, P.;
Schmidt, E. W.; Selsted, M. E.; Severinov, K.; Shen, B.; Sivonen, K.;
Smith, L.; Stein, T.; Sussmuth, R. E.; Tagg, J. R.; Tang, G.-L.; Vederas,
J. C.; Walsh, C. T.; Walton, J. D.; Willey, J. M.; van der Donk, W. A.
Nat. Prod. Rep. 2013, 30, 108.
̈
(34) Heal, W. P.; Wright, M. H.; Thinon, E.; Tate, E. W. Nat. Protoc.
2012, 7, 105.
(35) Watanabe, T.; Miyata, Y.; Abe, R.; Muranaka, N.; Hohsaka, T.
ChemBioChem 2008, 9, 1235.
(2) Knerr, P. J.; van der Donk, W. A. Annu. Rev. Biochem. 2012, 81,
479.
(36) Scheck, R. A.; Francis, M. B. ACS Chem. Biol. 2007, 2, 247.
(37) Rabuka, D.; Rush, J. S.; deHart, G. W.; Wu, P.; Bertozzi, C. R.
Nat. Protoc. 2012, 7, 1052.
(38) Cohen, J. D.; Zou, P.; Ting, A. Y. ChemBioChem 2012, 13, 888.
(39) Agarwal, P.; van der Weijden, J.; Sletten, E. M.; Rabuka, D.;
Bertozzi, C. R. Proc. Natl. Acad. Sci. U. S. A. 2013, 110, 46.
(40) Kellner, R.; Jung, G.; Josten, M.; Kaletta, C.; Entian, K. D.; Sahl,
H. G. Angew. Chem. 1989, 101, 618.
(3) Piper, C.; Cotter, P. D.; Ross, R. P.; Hill, C. Curr. Drug Discovery
Technol. 2009, 6, 1.
(4) Hasper, H. E.; Kramer, N. E.; Smith, J. L.; Hillman, J. D.;
Zachariah, C.; Kuipers, O. P.; de Kruijff, B.; Breukink, E. Science 2006,
313, 1636.
(5) Zhao, M.; Li, Z.; Bugenhagen, S. J. Nucl. Med. 2008, 49, 1345.
(6) Okuda, K.; Aso, Y.; Nakayama, J.; Sonomoto, K. J. Bacteriol. 2008,
190, 356.
(41) Holo, H.; Jeknic, Z.; Daeschel, M.; Stevanovic, S.; Nes, I. F.
Microbiology 2001, 147, 643.
(7) Schoof, S.; Baumann, S.; Ellinger, B.; Arndt, H. D. ChemBioChem
2009, 10, 242.
(42) Li, B.; Sher, D.; Kelly, L.; Shi, Y.; Huang, K.; Knerr, P. J.;
Joewono, I.; Rusch, D.; Chisholm, S. W.; van der Donk, W. A. Proc.
Natl. Acad. Sci. U.S.A. 2010, 107, 10430.
(8) Ross, A. C.; Liu, H.; Pattabiraman, V. R.; Vederas, J. C. J. Am.
Chem. Soc. 2010, 132, 462.
(9) Liu, W.; Chan, A. S.; Liu, H.; Cochrane, S. A.; Vederas, J. C. J.
Am. Chem. Soc. 2011, 133, 14216.
(43) Tang, W.; van der Donk, W. A. Biochemistry 2012, 51, 4271.
10370
dx.doi.org/10.1021/ja4010706 | J. Am. Chem. Soc. 2013, 135, 10362−10371