Ö. Dilek, S. L. Bane / Bioorg. Med. Chem. Lett. 19 (2009) 6911–6913
6913
equipment. The synthetic ease by which these molecules can be
prepared from the starting material should make these molecules
accessible to many laboratories that normally lack synthetic
capabilities.
0.6
0.5
0.4
0.3
0.2
0.1
0.0
Acknowledgments
We are thankful to Dr. Jürgen Schulte for collecting us the 11B-
NMR spectra. Thanks to David Tuttle for photography. Support for
this project was provided by National Institute of Health (R01
CA69571) and by the Research Foundation of the State University
of New York.
10x
Supplementary data
500
550
600
Supplementary data associated with this article can be found, in
Wavelength (nm)
Figure 4. Absorption spectra of 2 and 3 in methanol and pH 7 buffer. Samples were
prepared in the following manner: An aliquot of a stock solution in methanol was
removed and the solvent was evaporated. Methanol or 10 mM phosphate buffer, pH
References and notes
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Figure 5. SDS–PAGE of bovine brain tubulin labeled with compound
visualized using long wavelength UV light from a hand held device. Lane 1—10:1
dye:protein; Lane 2—20:1 dye:protein; Lane 3—40:1 dye:protein. The upper band is
5 and
a-tubulin and the lower band is b-tubulin.
pound 5 using standard lysine labeling conditions.26 Figure 5
shows a concentration dependent increase in fluorescence of both
the a- and b-subunits of tubulin. Even though there are two reac-
tive groups per fluorophore, no evidence of protein crosslinking
was detected in the gel; that is, no bands of higher molecular mass
were detected by either protein staining or fluorescence visualiza-
tion (data not shown). It was therefore unnecessary to use the
compound with a single chemically reactive succidimidyl ester
(6) for protein labeling. The fluorescent signal appears to be stable:
the photograph in Figure 5 was taken after the gels were stored
overnight in acetic acid:methanol:water destaining solution.
In summary, a 3,5-disubstituted BDP dye with strong absor-
bance near 543 nm was derivatized to extend the biological appli-
cations of molecules in this series. The dicarboxylic acid 3 retains
the desirable spectral properties of the thioether 2 while possess-
ing high aqueous solubility. Compound 3 can also be converted
to a disuccidimidyl ester that can be used for general protein label-
ing. The reactions to prepare compounds 3, 5, and 7 are straightfor-
ward and can be accomplished with very little specialized