Current Chemical Genomics and Translational Medicine
2008, 2 : 48-50Published online 2008 November 14. DOI: 10.2174/1875397300802010048
Publisher ID: CCGTM-2-48
RESEARCH ARTICLE
A Novel Fluorogenic Coumarin Substrate for Monitoring Acid Phosphatase Activity at Low pH Environment
1
Department of Chemistry & Chemical Engineering, Baoji University of Arts & Science, Shaanxi, China
2 Northwest University, Department of Chemistry, 229 North Taibei Road, Xian, Shaanxi, China
3 ABD Bioquest, Inc., 923 Thompson Place, Sunnyvale, CA 94085, USA
* Address correspondence to this author at the ABD Bioquest, Inc., 923 Thompson Place, Sunnyvale, CA 94085, USA; E-mail: jack@abdbioquest.com
2 Northwest University, Department of Chemistry, 229 North Taibei Road, Xian, Shaanxi, China
3 ABD Bioquest, Inc., 923 Thompson Place, Sunnyvale, CA 94085, USA
* Address correspondence to this author at the ABD Bioquest, Inc., 923 Thompson Place, Sunnyvale, CA 94085, USA; E-mail: jack@abdbioquest.com
ABSTRACT
This article described the synthesis and application of 6-chloro-8-fluoro-4-methylumbelliferone phosphate (CF-MUP) in analyzing acid phosphatase activity. Compared to the existing MUP, the new coumarin phosphate, CF-MUP, demonstrateed much higher sensitivity and was more robust for detecting the activity of acid phosphatase than the classic substrate 4-methylumbelliferone phosphate (MUP). The product of enzyme reaction, 6-chloro-8-fluoro-4-methylumbelliferone (CF-MU) possesses strong fluorescence at ~450 nm with low pKa (4.7), high fluorescence quantum yield and pH independence in the physiological pH range. This new fluorescence dye, CF-MU, is a convenient tool for assays with buffer pH between 4.5 and 8.