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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Robison, Aaron D. Cremer, Paul S. Huang, Da Liu, Yiquan |
| Description | Country affiliation: United States Author Affiliation: Huang D ( Department of Chemistry, Texas A&M University, 3255 TAMU, College Station, TX 77843, USA.) |
| Abstract | We have developed a technique for sensing protein-small molecule and protein-ion interactions in bulk aqueous solution by utilizing a pH sensitive dye, 5-(and-6)-carboxyfluorescein, conjugated to free lysine residues on the surfaces of designated capture proteins. The fluorescein intensity was found to change by about 6% and 15% for small molecule and ion binding, respectively. The assay works by modulating the local electric fields around a pH sensitive dye. This, in turn, alters the dye's apparent pK(A) value. Such changes may result directly from the charge on the analyte, occur through allosteric effects related to the binding process, or result from a combination of both. The assay was used to follow the binding of Ca(2+) to calmodulin (CaM) and thiamine monophosphate (ThMP) to thiamine binding protein A (TbpA). The results demonstrate a binding constant of 1.1 µM for the Ca(2+)/CaM pair and 3.2 nM for ThMP/TbpA pair, which are in excellent agreement with literature values. These assays demonstrate the generality of this method for observing the interactions of small molecules and ions with capture proteins. In fact, the assay should work as a biosensor platform for most proteins containing a specific ligand binding site, which would be useful as a simple and rapid preliminary screen of protein-ligand interactions. |
| ISSN | 09565663 |
| Issue Number | 1 |
| Volume Number | 38 |
| e-ISSN | 18734235 |
| Journal | Biosensors and Bioelectronics |
| Language | English |
| Publisher | Elsevier |
| Publisher Date | 2012-10-01 |
| Publisher Place | Great Britain (UK) |
| Access Restriction | One Nation One Subscription (ONOS) |
| Subject Keyword | Calmodulin Metabolism Escherichia Coli Proteins Escherichia Coli Fluoresceins Fluorescent Dyes Animals Binding Sites Biosensing Techniques Methods Calcium Cattle Chemistry Hydrogen-ion Concentration Ions Ligands Protein Binding Spectrometry, Fluorescence Thiamine Titrimetry Journal Article Research Support, U.s. Gov't, Non-p.h.s. Discipline Biotechnology |
| Content Type | Text |
| Resource Type | Article |
| Subject | Nanoscience and Nanotechnology Medicine Biophysics Biomedical Engineering Biotechnology Electrochemistry |
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