Y. Ambroise et al. / Bioorg. Med. Chem. Lett. 10 (2000) 1125±1127
1127
Figure 1. UV-spectral recording of the photoirradiation of 1, 2 and 3 at 18 ꢀC. 100 mM of 1 or 2 or 3 in 50 mM Tris buer, 50 mM NaCl, pH=8. 1
l=250 nm, E=5.10 5 mW/cm2, 0, 10, 30, 60, 90 and 120 s. 2 l=300 nm, E=10 5 mW/cm 2, 0,30, 60, 90, 120, 150, 180, 210, and 240 sec. 3 l=366
nm, E=1.5 mW/cm 2, 0, 120, 300, 720, 1200, 2000 s.
These probes must have favorable photochemical prop-
erties if they are to be used as irreversible photochemical
markers. Figure 1 shows the absorption spectra of
probes 1, 2 and 3 as well as their photodecomposition
by irradiation in buered medium. Irradiation of each
compound led to its complete disappearance, and the
observed isobestics are illustrative of a unique photo-
decomposition process.
7. Pourcher, T.; Bibi, E.; Kaback, H. R.; Leblanc, G. Bio-
chemistry 1996, 35, 4161.
8. Bot®eld, M. C.; Naguchi, K.; Tsuchiya, T.; Wilson, T. H. J.
Biol. Chem. 1992, 267, 1818.
9. Gwizdek, C.; Leblanc, G.; Bassilana, M. Biochemistry 1997,
36, 8522.
10. Mus-Veteau, I.; Pourcher, T.; Leblanc, G. Biochemistry
1995, 34, 6775.
11. Maehrel, C.; Cordat, E.; Mus-Veteau, I.; Leblanc, G. J.
Biol. Chem. 1998, 273, 33192.
In summary, three new photoanity analogues and
their tritiated form have been prepared, in order to map
the binding site of melibiose permease. Two of these
photoactivatable compounds gave excellent preliminary
results with a high anity for the melibiose permease of
Escherichia coli. These probes should also be useful for
mapping the binding site of other sugar transporters
that have a substrate speci®city similar to that of MelB
(e.g., lactose permease). Photoanity labeling experi-
ments with MelB are currently underway and will be
reported elsewhere.23
12. Lacapere, J. J.; Stokes, D. L.; Mosser, G.; Rank, J. L.;
Leblanc, G.; Rigaud, J. L. Ann. N.Y. Acad. Sci. 1997, 834, 9.
13. Kotzyba-Hibert, F.; Kapfer, I.; Goeldner, M. Angew.
Chem., Int. Ed. Engl. 1995, 34, 1296.
14. Damiano-Forano, E.; Bassilana, M.; Leblanc, G. J. Biol.
Chem. 1986, 261, 6893.
15. Characterization data for compound 1: 1H NMR (300
MHz, D2O) d 3.75 (m, 2H), 3.94 (dd, J=2, 9 Hz, 1H), 4.12±
4.15 (m, 3H), 6.08 (d, J=2 Hz, 1H), 7.64 (d, J=9 Hz, 2H),
8.58 (d, J=9 Hz, 2H); 13C NMR (75 MHz, CDCl3) 61.3, 68.0,
69.3, 69.6, 73.4, 98.2, 104.0, 116.5, 120.2, 162.5, 167.3; MS
(electrospray) m/z 283 (M+); HPLC, Zorbax-ODS, H2O/
CF3COOH 100/0.1; VR=6.4 min.
16. Lamotte, H.; Degeilh, F.; Rousseau, B.; Neau, P.;
Ripoche, P. Bioorg. Med. Chem. Lett. 1993, 3, 2587.
17. Characterization data for compound 2: 1H NMR (300
MHz, D2O) d 3.90 (d, J=7 Hz, 2H), 4±410.28 (m, 4H), 5.80
(d, J=2 Hz, 1H), 7.27 (d, J=9 Hz, 2H), 8.38 (d, J=9 Hz, 2H);
13C NMR (75 MHz, D2O) 61.5, 68.6, 69.7, 69.9, 72.2, 98.4,
119.7, 120.6, 134.9, 153.9 MS (CI, NH3) m/z 315 (MNH+4 );
HPLC, Zorbax-ODS, H2O:CH3OH:CF3COOH 80/20/0.1;
VR=15.8 min.
Acknowledgements
We are grateful to Morgane Le Bastard, Raymonde
Lemonnier and Alain Valleix for skillful technical
assistance.
References and Notes
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20. Characterization data for compound 3: 1H NMR (300
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bax-ODS, H2O:CH3CN:CF3COOH 80:20:0.1; VR=18.3 min.
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