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The total synthesis of ramoplanin aglycone has recently been reported by Boger and co-workers. See: (a) Jiang, W.; Wanner, J.; Lee, R. J.; Bounaud, P. Y.; Boger, D. L. J. Am. Chem. Soc. 2002, 124, 5288-5290.
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It has recently been shown, however, that ramoplanin binds to Lipid I analogues. See: (a) Lo, M.-C. Ph.D. Thesis. Princeton University, 2000. (b) Cudic, P.; Kranz, J. K.; Behenna, D. C.; Kruger, R. G.; Tadesse, H.; Wand, A. J.; Veklich, Y. I.; Weisel, J. W.; McCafferty, D. G. Proc. Natl. Acad. Sci. U.S.A. 2002, 99, 7384-7389. (c) Cudic, P.: Behenna, D. C.; Kranz, J. K.; Kruger, R. G.; Wand, A. J.; Veklich, Y. I.; Weisel, J. W.; McCafferty, D. G. Chem. Biol. 2002, 9, 897-906.
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It has recently been shown, however, that ramoplanin binds to Lipid I analogues. See: (a) Lo, M.-C. Ph.D. Thesis. Princeton University, 2000. (b) Cudic, P.; Kranz, J. K.; Behenna, D. C.; Kruger, R. G.; Tadesse, H.; Wand, A. J.; Veklich, Y. I.; Weisel, J. W.; McCafferty, D. G. Proc. Natl. Acad. Sci. U.S.A. 2002, 99, 7384-7389. (c) Cudic, P.: Behenna, D. C.; Kranz, J. K.; Kruger, R. G.; Wand, A. J.; Veklich, Y. I.; Weisel, J. W.; McCafferty, D. G. Chem. Biol. 2002, 9, 897-906.
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Kranz, J.K.2
Behenna, D.C.3
Kruger, R.G.4
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Wand, A.J.6
Veklich, Y.I.7
Weisel, J.W.8
McCafferty, D.G.9
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12
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It has recently been shown, however, that ramoplanin binds to Lipid I analogues. See: (a) Lo, M.-C. Ph.D. Thesis. Princeton University, 2000. (b) Cudic, P.; Kranz, J. K.; Behenna, D. C.; Kruger, R. G.; Tadesse, H.; Wand, A. J.; Veklich, Y. I.; Weisel, J. W.; McCafferty, D. G. Proc. Natl. Acad. Sci. U.S.A. 2002, 99, 7384-7389. (c) Cudic, P.: Behenna, D. C.; Kranz, J. K.; Kruger, R. G.; Wand, A. J.; Veklich, Y. I.; Weisel, J. W.; McCafferty, D. G. Chem. Biol. 2002, 9, 897-906.
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Enzyme Kinetics
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Several studies indicate that ramoplanin recognizes the pyrophosphate portion of the substrate. See ref 4. This suggests that electrostatic interactions with one or both ornithines could be important for binding. A recent NMR structure of a ramoplanin dimer shows that the ornithine amines flank separate clefts, suggesting that only one of them can contact the pyrophosphate. See: Lo, M. C.; Helm, J. S.; Samgadharan, G.; Pelczer, I.; Walker, S. J. Am. Chem. Soc. 2001, 123, 8640-8641. Therefore, we reasoned that acylating with alanine might eliminate a specific interaction with the substrate while maintaining the net charge on the molecule. The dialanine derivative was also synthesized and behaved similarly to compound 3.
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The binding isotherm was fit by nonlinear least-squares regression. See: Reid, S. L.; Parry, D.; Liu, H.-H.; Connolly, B. A. Biochemistry 2001, 40, 2484-2494.
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MICs were measured against E. faecium strain L19624 and E. faecalis strain Z9212 according to standard methods. See: Methods for Dilution Anti Microbial Susceptibility Tests for Bacteria that Grow Aerobically (approved standard, NCCLS Document M7-A4, National Committee for Clinical Laboratory Standards, Wayne, PA, ed. 4, 1997).
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