| 光谱学与光谱分析 |
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| Kinetic Study on Inhibition Effects of Dansyl-L-Phenylalanine and L-Phenylalanine on Calf Intestinal Alkaline phosphatase |
| LI Li-na1, 2, WU Yu-qing1*, René Buchet2 |
1. State Key Laboratory for Supramolecular Structure and Materials, Jilin University, Changchun 130012, China
2. Université de Lyon, Villeurbanne, F-69622, France; CNRS UMR 5246 ICBMS, Villeurbanne, F-69622, France |
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Abstract To evaluate the inhibition effect of dansyl-L-phenylalanine on calf intestinal alkaline phosphatase (CIAP), UV-Vis spectrophotometric method was employed. It was found that dansyl-L-phenylalanine can selectively inhibit CIAP. The kinetic inhibition processes of dansyl-L-phenylalanine and L-phenylalanine were comparatively studied. The authors’ finding elucidates that at the optimized alkaline pH of alkaline phosphatase (pH 10.4) and 37 ℃, dansyl-L-phenylalanine can inhibit alkaline phosphatase activity of CIAP efficiently and specifically, similar as L-phenylalanine. Both inhibition types were uncompetitive inhibition resulting from the double reciprocal curve fitting of v versus substrate concentrations, and the inhibition constants Ki of both inhibitors were determined to be 2.3 and 1.1 mmol·L-1 respectively, both of which were at millimolar level. The investigation of the inhibition effect of dansyl modified L-phenylalanine on calf intestinal alkaline phosphatase not only helped get insight into the detailed inhibition mechanism of L-phenylalanine on tissue specific alkaline phosphatase, such as in the case of intestinal alkaline phosphatase, but also provided the possibility to employ fluorescence spectroscopy by labeling the specific inhibitors of alkaline phosphatase with chromophoric groups.
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Received: 2008-08-10
Accepted: 2008-12-20
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Corresponding Authors:
WU Yu-qing
E-mail: yqwu@jlu.edu.cn
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