New method for covalent fluorescent biomolecule labeling with hemicyanine dye.
Fluorescent chromophore, alkylamino-(tetra-hydronaphthalenylidene)- benzothiazolium derivatives (HBTN dyes), are proposed as covalent labels for proteins via aliphatic amino groups. Spectral-luminescent properties of 3-methyl-2-{(E)-[7-(methylamino)-4,4a,5,6-tetra-hydronaphthalen-2(3H)-ylidene]methyl}-1,3-benzothiazol-3-ium chloride (HBTN, R=Me) and its predecessor, 2-[(E)-(7-methoxy-4,4a,5,6-tetrahydronaphthalen-2(3H)-ylidene)methyl]-3-methyl-1,3-benzothiazol-3-ium chloride (ABTN), are studied for free dyes and in the presence of DNA and BSA. Considerable spectral-luminescent changes accompany the transformation of ABTN into HBTN that allows monitoring conjugation reaction. In presence of DNA and BSA the HBTN increases its emission in 15 and 4 times respectively and becomes strongly fluorescent. The conditions for labeling are developed and a model conjugate of HBTN dye with BSA is synthesized. It was shown that using of HBTN dye as a fluorescent label allows detection by eye of about 3 mug/band of BSA on polyacrylamide gel upon UV-irradiation.
Kostenko
Olexander M
OM
Institute of Molecular Biology and Genetics, National Academy of Sciences of Ukraine, 150 Zabolotnogo St., 03143 Kyiv, Ukraine.
Kovalska
Vladyslava B
VB
Volkova
Kateryna D
KD
Shaytanov
Pavel
P
Kocheshev
Igor O
IO
Slominskiy
Yuriy L
YL
Pisareva
Irina V
IV
Yarmoluk
Sergiy M
SM
D000818:Animals; D002417:Cattle; D002853:Chromatography, Liquid; D004247:DNA / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D005456:Fluorescent Dyes / Q000138:chemical synthesis / Q000737:chemistry*; D013058:Mass Spectrometry; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000378:metabolism* / Q000528:radiation effects; D013050:Spectrometry, Fluorescence / Q000379:methods*; D014466:Ultraviolet Rays
D005456:Fluorescent Dyes; D011506:Proteins; D012710:Serum Albumin, Bovine; D004247:DNA