Synthesis of fluorescent-labeled aeruginosin derivatives for high-throughput fluorescence correlation spectroscopy assays

• Published in: Bioorganic & medicinal chemistry letters Vol. 17; no. 10; pp. 2904 - 2907
• Main Authors: Hoshina, Yoichiro, Yamada, Yoshifumi, Tanaka, Hiroshi, Doi, Takayuki, Takahashi, Takashi
• Format: Journal Article
• Language: English
• Published: Oxford Elsevier Ltd 15.05.2007; Elsevier
• Subjects: Analytical, structural and metabolic biochemistry; Biological and medical sciences; Chemical biology; Combinatorial chemistry; Enzymes and enzyme inhibitors; Fluorescence correlation spectroscopy; Fluorescent Dyes; Fundamental and applied biological sciences. Psychology; Hydrolases; Leucine - analogs & derivatives; Leucine - chemical synthesis; Leucine - chemistry; Medical sciences; Miscellaneous; Molecular Structure; Pharmacology. Drug treatments; Solid-phase synthesis; Spectrometry, Fluorescence - methods; Spectrophotometry, Ultraviolet; Chemical biology; Combinatorial chemistry; Solid-phase synthesis; Fluorescence correlation spectroscopy; Serine endopeptidases; Enzyme; Enzyme inhibitor; Guanidines; Basic dye; In vitro; Fluorescence spectrometry; Peptidases; Competitive fixation; Trypsin; Structure activity relation; Fluorescent labelling; Chemical compound library; Hydrolases; Solid phase; Chemical synthesis; Xanthene dye
• ISSN: 0960-894X; 1464-3405
• DOI: 10.1016/j.bmcl.2007.02.045

The design and solid-phase synthesis of effective fluorescent-labeled aeruginosin derivatives and their application to the fluorescence correlation spectroscopy (FCS)-based competitive binding assay of an aeruginosin library are described. The phenolic hydroxyl group on the ( R)-3-(4-hydroxyphenyl)lactic acid ( d-Hpla) residue was observed to be suitable for connecting Rhodamine green derivative with minimum loss of biological activity. In addition, the FCS-based binding assay of the library using fluorescent-labeled chemical probes was also achieved.