High-density miniaturized thermal shift assays as a general strategy for drug discovery.
AuthorsPantoliano MW, Petrella EC, Kwasnoski JD, Lobanov VS, Myslik J, Graf E, Carver T, Asel E, Springer BA, Lane P, Salemme FR
JournalJ Biomol Screen
PubMed ID11788061
'More general and universally applicable drug discovery assay technologies are needed in order to keep pace with the recent advances in combinatorial chemistry and genomics-based target generation. Ligand-induced conformational stabilization of proteins is a well-understood phenomenon in which substrates, inhibitors, cofactors, and even other proteins provide enhanced stability to proteins ... More
Label-free continuous enzyme assays with macrocycle-fluorescent dye complexes.
AuthorsHennig A, Bakirci H, Nau WM
JournalNat Methods
PubMed ID17603491
We introduce a new economic, convenient and general assay principle based on the reversible interaction of water-soluble macrocycles and fluorescent dyes. We show that amino acid decarboxylase activity can be continuously monitored by measuring changes in fluorescence, which result from the competition of the enzymatic product and the dye for ... More
The use of differential scanning fluorimetry to detect ligand interactions that promote protein stability.
AuthorsNiesen FH, Berglund H, Vedadi M,
JournalNat Protoc
PubMed ID17853878
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 ... More
Decrypting the biochemical function of an essential gene from Streptococcus pneumoniae using ThermoFluor technology.
The protein product of an essential gene of unknown function from Streptococcus pneumoniae was expressed and purified for screening in the ThermoFluor affinity screening assay. This assay can detect ligand binding to proteins of unknown function. The recombinant protein was found to be in a dimeric, native-like folded state and ... More