A Novel Fluorogenic Coumarin Substrate for Monitoring Acid Phosphatase Activity at Low pH Environment
Desuo Yang1, Zongxiao Li1, Yubo Allan Diwu3, Hanzhuo Fu2, Jinfang Liao3, Chunmei Wei3, Zhenjun Diwu*, 3
Identifiers and Pagination:Year: 2008
First Page: 48
Last Page: 50
Publisher Id: CCGTM-2-48
Article History:Received Date: 1/9/2008
Revision Received Date: 17/10/2008
Acceptance Date: 20/10/2008
Electronic publication date: 14/11/2008
Collection year: 2008
open-access license: This is an open access article licensed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted, non-commercial use, distribution and reproduction in any medium, provided the work is properly cited.
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.