The conversion of azo-quenchers to fluorophores

O Hofstetter; H Hofstetter; T Miron; M Wilchek

doi:10.1016/j.ab.2019.113400

The conversion of azo-quenchers to fluorophores

Anal Biochem. 2019 Nov 15:585:113400. doi: 10.1016/j.ab.2019.113400. Epub 2019 Aug 19.

Authors

O Hofstetter¹, H Hofstetter², T Miron³, M Wilchek⁴

Affiliations

¹ Department of Chemistry and Biochemistry, Northern Illinois University, DeKalb, IL, 60115, USA.
² Department of Chemistry, University of Wisconsin-Madison, Madison, WI, 53706, USA.
³ Department of Biomolecular Science, Weizmann institute of Science, Rehovot, Israel. Electronic address: talia.miron@weizmann.ac.il.
⁴ Department of Biomolecular Science, Weizmann institute of Science, Rehovot, Israel.

PMID: 31437428
DOI: 10.1016/j.ab.2019.113400

Abstract

In this short note we describe the conversion of the widely used fluorescence quenching azo-dyes DABCYL and HABA to fluorophores. The dyes were conjugated to the proteins RNase and human serum albumin (HSA) and subsequently reduced using sodium dithionite (Na₂S₂O₄), thus forming amine-containing fluorophores. Since this chemical reaction can be applied to any azo-containing quencher compound, a great variety of substances can be readily obtained synthetically. This approach provides a promising tool in the use of fluorescence-based investigations of biomolecular interactions.

Keywords: Azo dye; Beacon; FRET; Fluorophore; Quencher.

MeSH terms

Amino Acid Sequence
Azo Compounds / chemistry*
Binding Sites
Dithionite / chemistry
Fluorescence Resonance Energy Transfer
Fluorescent Dyes / chemistry*
Humans
Molecular Structure
Oxidation-Reduction
Protein Binding
Ribonucleases / chemistry
Serum Albumin / chemistry
Structure-Activity Relationship
p-Dimethylaminoazobenzene / analogs & derivatives*
p-Dimethylaminoazobenzene / chemistry

Substances

Azo Compounds
Fluorescent Dyes
Serum Albumin
Dithionite
HABA
4-(4-dimethylaminophenylazo)benzoic acid
p-Dimethylaminoazobenzene
Ribonucleases