RESUMO
The group II azoreductase BTI1 utilizes NADPH to directly cleave azo bonds in water-soluble azo dyes, including quenchers of fluorescence. Unexpectedly, optimal reduction was dye specific, ranging from a pH of <5.5 for Janus green B, to pH 6.0 for methyl red, methyl orange, and BHQ-10, to pH >8.3 for flame orange.
Assuntos
Compostos Azo/metabolismo , NADH NADPH Oxirredutases/metabolismo , NADP/metabolismo , Concentração de Íons de Hidrogênio , Nitrorredutases , Especificidade por SubstratoRESUMO
Cyanine and squaraine dyes that contain one or two phosphonate groups attached directly to the aromatic residues of the dyes were synthesized. Absorption and fluorescence properties of the dyes were determined. Succinimidyl active esters were prepared from the dyes and were used to label proteins and oligonucleotides. Some of the dyes permit the preparation of brighter conjugates than do commercially available analogues such as Cy3 and Cy5.
