Biotinylated indoles as probes for indole-binding proteins

被引:13
作者
Dolusic, E
Kowalczyk, M
Magnus, V
Sandberg, G
Normanly, J
机构
[1] Rudjer Boskovic Inst, Dept Mol Genet, HR-10002 Zagreb, Croatia
[2] Swedish Univ Agr Sci, Dept Forest Genet & Plant Physiol, S-90183 Umea, Sweden
[3] Univ Massachusetts, Dept Biochem & Mol Biol, Amherst, MA 01003 USA
关键词
D O I
10.1021/bc000035o
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Biotinylated indoles were prepared for application as bifunctional probes for the detection of indole-binding proteins which participate in the life processes of humans, animals, plants, and bacteria. The indole nucleus was functionalized, at ring positions 3, 5, or 6, by attachment of a 2-aminoethyl group, which was then coupled to the carboxyl moiety of biotin, via a spacer composed of 3 or 4 concatenated beta -alanine residues. The constructs thus obtained were able to inhibit tryptophanase activity, similarly to indole in a concentration-dependent manner. They also bound strongly to lysozyme and weakly to bovine and human serum albumins, in accordance with the known affinities of these proteins for indole and 3-(2-aminoethyl)indole (tryptamine). The biotin end of the protein-bound bifunctional probes could then be detected by coupling to (strept)avidin conjugated to alkaline phosphatase or horseradish peroxidase, followed by incubation with substrates which are converted by these enzymes to intensely colored or chemiluminescent products.
引用
收藏
页码:152 / 162
页数:11
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共 50 条
[21]   INDOLE OXYGENASE FROM THE LEAVES OF TECOMA STANS [J].
KUNAPULI, SP ;
VAIDYANATHAN, CS .
PLANT SCIENCE LETTERS, 1982, 24 (02) :183-188
[22]   3-DIMENSIONAL STRUCTURES OF AVIDIN AND THE AVIDIN-BIOTIN COMPLEX [J].
LIVNAH, O ;
BAYER, EA ;
WILCHEK, M ;
SUSSMAN, JL .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1993, 90 (11) :5076-5080
[23]  
MCCORMICK D. B., 1970, Methods in Enzymology, V18A, P383, DOI 10.1016/0076-6879(71)18331-0
[24]   ASSOCIATION OF INDOLE ANALOGUES TO DEFATTED HUMAN SERUM ALBUMIN [J].
MCMENAMY, RH .
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1963, 103 (03) :409-&
[25]  
MCMENAMY RH, 1963, J BIOL CHEM, V238, P3241
[26]   A deletion in an indole synthase gene is responsible for the DIMBOA-deficient phenotype of bxbx maize [J].
Melanson, D ;
Chilton, MD ;
MastersMoore, D ;
Chilton, WS .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1997, 94 (24) :13345-13350
[27]  
MORINO Y, 1967, J BIOL CHEM, V242, P2793
[28]   INDOLE OXIDASE ISOLATED FROM LEAVES OF TECOMA STANS [J].
NAIR, PM ;
VAIDYANATHAN, CS .
BIOCHIMICA ET BIOPHYSICA ACTA, 1964, 81 (03) :496-&
[29]   PROGRESSION OF GLOMERULAR SCLEROSIS IN EXPERIMENTAL UREMIC RATS BY ADMINISTRATION OF INDOLE, A PRECURSOR OF INDOXYL SULFATE [J].
NIWA, T ;
ISE, M ;
MIYAZAKI, T .
AMERICAN JOURNAL OF NEPHROLOGY, 1994, 14 (03) :207-212
[30]   RETHINKING AUXIN BIOSYNTHESIS AND METABOLISM [J].
NORMANLY, J ;
SLOVIN, JP ;
COHEN, JD .
PLANT PHYSIOLOGY, 1995, 107 (02) :323-329