R. G. Kruger et al. / Bioorg. Med. Chem. 12 (2004) 3723–3729
3729
3. Mazmanian, S. K.; Liu, G.; Jensen, E. R.; Lenoy, E.;
Schneewind, O. Proc. Natl. Acad. Sci. U.S.A. 2000, 97(10),
5510.
4. Bolken, T. C.; Franke, C. A.; Jones, K. F.; Zeller, G. O.;
Jones, C. H.; Dutton, E. K.; Hruby, D. E. Infect. Immun.
2001, 69(1), 75.
5. Garandeau, C.; Reglier-Poupet, H.; Dubail, I.; Beretti,
J. L.; Berche, P.; Charbit, A. Infect. Immun. 2002, 70(3), 1382.
6. Osaki, M.; Takamatsu, D.; Shimoji, Y.; Sekizaki, T.
J. Bacteriol. 2002, 184(4), 971.
7. Kharat, A. S.; Tomasz, A. Infect. Immun. 2003, 71(5), 2758.
8. Lee, S. F.; Boran, T. L. Infect. Immun. 2003, 71(2), 676.
9. Barnett, T. C.; Scott, J. R. J. Bacteriol. 2002, 184(8), 2181.
10. Kim, S. W.; Chang, I. M.; Oh, K. B. Biosci., Biotechnol.,
Biochem. 2002, 66(12), 2751.
11. Kim, S. H.; Shin, D. S.; Oh, M. N.; Chung, S. C.; Lee,
J. S.; Chang, I. M.; Oh, K. B. Biosci., Biotechnol.,
Biochem. 2003, 67(11), 2477.
12. Kim, S. H.; Shin, D. S.; Oh, M. N.; Chung, S. C.; Lee,
J. S.; Oh, K. B. Biosci., Biotechnol., Biochem. 2004, 68(2),
421.
13. Scott, C. J.; McDowell, A.; Martin, S. L.; Lynas, J. F.;
Vandenbroeck, K.; Walker, B. Biochem. J. 2002, 366(Pt 3),
953.
14. Palmer, J. T.; Rasnick, D.; Klaus, J. L.; Bromme, D.
J. Med. Chem. 1995, 38(17), 3193.
were coupled by automated procedures. The peptide was
cleaved from the resin using TFA–water (95:5) for 3 h.
Excess TFA was removed by rotary evaporation, the
peptide was precipitated using cold diethyl ether, filtered
with a fine porosity fritted glass filter, dissolved in water,
and lyophilized to afford the desired crude product 1,
NH2-YALPE-AlaW{PO2H–CH2}Gly-EE-NH2 (75 mg,
15%). The crude product was purified by HPLC using a
semipreparative C18 Jupiter column to P98% purity
and verified by MALDI-TOF MS.
4.4. Kinetic characterization of the NH2-YALPE-
AlaW{PO2H–CH2}Gly-EE-NH2 inhibitor
NH2-YALPE-AlaW{PO2H–CH2}Gly-EE-NH2 inhibi-
tory patterns were determined with 1 mM Abz-LPETG-
Dap(Dnp) and Gly5 concentrations from 50 to 1000 lM
using 100 nM SrtADN24. The enzyme was assayed using
the HPLC based method23 monitoring the formation of
the G-Dap(Dnp) product at 355 nm. The data were fit to
the Ping-Pong Bi Bi hydrolytic shunt kinetic mechanism
(Eq. 3) using the Grafit global fitting program (Eritha-
cus Software).
15. Connolly, K. M.; Smith, B. T.; Pilpa, R.; Ilangovan, U.;
Jung, M. E.; Clubb, R. T. J. Biol. Chem. 2003, 24, 24.
16. Frankel, B. A.; Bentley, M.; Kruger, R. G.; McCafferty,
D. G., J. Am. Chem. Soc. 2004, ASAP on the Web,
February 27.
Note added in proof
During the final stages of publication of this manuscript,
Oh and coworkers [J. Med. Chem. 2004, 47, 2418–2421]
recently reported the discovery of reversible inhibition
of SrtA by small molecule diarylacrylonitriles from
random library screening efforts. In this report, the
primary structure–activity relationship and minimal
structural requirements for potency were established
through structural modifications and modeling studies.
17. Copie, V.; Kolbert, A. C.; Drewry, D. H.; Bartlett, P. A.;
Oas, T. G.; Griffin, R. G. Biochemistry 1990, 29(39), 9176.
18. Allen, M. C.; Fuhrer, W.; Tuck, B.; Wade, R.; Wood,
J. M. J. Med. Chem. 1989, 32(7), 1652.
19. Yiallouros, I.; Vassiliou, S.; Yiotakis, A.; Zwilling, R.;
Stocker, W.; Dive, V. Biochem. J. 1998, 331(Pt 2), 375.
20. Buchardt, J.; Meldal, M. J. Chem. Soc., Perkin Trans. 1
2000, 3306.
21. Kruger, R. G.; Otvos, B.; Frankel, B. A.; Bentley, M.;
Dostal, P.; McCafferty, D. G. Biochemistry 2004, 43(6),
1541.
22. Baylis, E. K.; Campbell, C. D.; Dingwall, J. G. J. Chem.
Soc., Perkin Trans. 1 1984, 2845.
Acknowledgements
23. Kruger, R. G.; Dostal, P.; McCafferty, D. G. Anal.
Biochem. 2004, 326(1), 42.
24. Ton-That, H.; Liu, G.; Mazmanian, S. K.; Faull, K. F.;
Schneewind, O. Proc. Natl. Acad. Sci. U.S.A. 1999, 96(22),
12424.
We thank J. Drury for assistance in deriving rate
equations. This research was generously supported by
NIH Grants AI46611 and GM65539. B. A. Frankel is
the recipient of an NSF Predoctoral Fellowship.
25. Ton-That, H.; Mazmanian, S. K.; Alksne, L.; Schneewind,
O. J. Biol. Chem. 2002, 277(9), 7447.
26. Ton-That, H.; Mazmanian, S. K.; Faull, K. F.; Schnee-
wind, O. J. Biol. Chem. 2000, 275(13), 9876.
27. Huang, X.; Aulabaugh, A.; Ding, W.; Kapoor, B.; Alksne,
L.; Tabei, K.; Ellestad, G. Biochemistry 2003, 42(38),
11307.
References and notes
1. Mazmanian, S. K.; Liu, G.; Ton-That, H.; Schneewind, O.
Science 1999, 285(5428), 760.
2. Mazmanian, S. K.; Ton-That, H.; Su, K.; Schneewind, O.
Proc. Natl. Acad. Sci. U.S.A. 2002, 99(4), 2293.
28. Karkowsky, A. M.; Bergamini, M. V.; Orlowski, M.
J. Biol. Chem. 1976, 251(15), 4736.