title: Biotinylated indoles as probes for indole-binding proteins.
abstract: 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.
authors: Department of Molecular Genetics, Ruder Bosković Institute, p.p. 180, HR-10002 Zagreb, Croatia.
authors: E
authors: E
authors: Dolusić
authors: M
authors: M
authors: Kowalczyk
authors: V
authors: V
authors: Magnus
authors: G
authors: G
authors: Sandberg
authors: J
authors: J
authors: Normanly
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D001360:Avidin / Q000378:metabolism; D001710:Biotin / Q000737:chemistry; D019921:Biotinylation; D006801:Humans; D007211:Indoles / Q000737:chemistry* / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D015335:Molecular Probes / Q000737:chemistry*; D015394:Molecular Structure; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism; D012694:Serine / Q000378:metabolism; D012709:Serum Albumin / Q000378:metabolism; D014368:Tryptophanase / Q000378:metabolism
chemical_list: D007211:Indoles; D015335:Molecular Probes; D011506:Proteins; D012709:Serum Albumin; D001360:Avidin; D012694:Serine; D001710:Biotin; D000469:Alkaline Phosphatase; D014368:Tryptophanase