The use of differential scanning fluorimetry to detect ligand interactions that promote protein stability Export

@article{citeulike:4035947, abstract = {Differential scanning fluorimetry (DSF) is a rapid and inexpensive screening method to identify low-molecular-weight ligands that bind and stabilize purified proteins. The temperature at which a protein unfolds is measured by an increase in the fluorescence of a dye with affinity for hydrophobic parts of the protein, which are exposed as the protein unfolds. A simple fitting procedure allows quick calculation of the transition midpoint; the difference in the temperature of this midpoint in the presence and absence of ligand is related to the binding affinity of the small molecule, which can be a low-molecular-weight compound, a peptide or a nucleic acid. DSF is best performed using a conventional real-time PCR instrument. Ligand solutions from a storage plate are added to a solution of protein and dye, distributed into the wells of the PCR plate and fluorescence intensity measured as the temperature is raised gradually. Results can be obtained in a single day.}, author = {Niesen, Frank H. and Berglund, Helena and Vedadi, Masoud}, citeulike-article-id = {4035947}, citeulike-linkout-0 = {http://dx.doi.org/10.1038/nprot.2007.321}, citeulike-linkout-1 = {http://dx.doi.org/10.1038/nprot.2007.321}, citeulike-linkout-2 = {http://view.ncbi.nlm.nih.gov/pubmed/17853878}, citeulike-linkout-3 = {http://www.hubmed.org/display.cgi?uids=17853878}, doi = {10.1038/nprot.2007.321}, issn = {1750-2799}, journal = {Nat. Protocols}, keywords = {dsf, inhibitor}, month = {September}, number = {9}, pages = {2212--2221}, posted-at = {2009-02-11 16:18:17}, priority = {2}, title = {The use of differential scanning fluorimetry to detect ligand interactions that promote protein stability}, url = {http://dx.doi.org/10.1038/nprot.2007.321}, volume = {2}, year = {2007} }

TY - JOUR ID - citeulike:4035947 L3 - citeulike-article-id:4035947 N2 - Differential scanning fluorimetry (DSF) is a rapid and inexpensive screening method to identify low-molecular-weight ligands that bind and stabilize purified proteins. The temperature at which a protein unfolds is measured by an increase in the fluorescence of a dye with affinity for hydrophobic parts of the protein, which are exposed as the protein unfolds. A simple fitting procedure allows quick calculation of the transition midpoint; the difference in the temperature of this midpoint in the presence and absence of ligand is related to the binding affinity of the small molecule, which can be a low-molecular-weight compound, a peptide or a nucleic acid. DSF is best performed using a conventional real-time PCR instrument. Ligand solutions from a storage plate are added to a solution of protein and dye, distributed into the wells of the PCR plate and fluorescence intensity measured as the temperature is raised gradually. Results can be obtained in a single day. IS - 9 JF - Nat. Protocols SN - 1750-2799 EP - 2221 TI - The use of differential scanning fluorimetry to detect ligand interactions that promote protein stability VL - 2 SP - 2212 KW - dsf KW - inhibitor AU - Niesen, Frank AU - Berglund, Helena AU - Vedadi, Masoud PY - 2007/09// UR - http://dx.doi.org/10.1038/nprot.2007.321 ER -