Open AccessEnthalpy arrays
Author(s) -
Francisco Eduardo Torres,
Peter Kühn,
Dirk De Bruyker,
Alan G. Bell,
Michal V. Wolkin,
Eric Peeters,
James R. Williamson,
G. B. Anderson,
Gregory P. Schmitz,
Michael I. Recht,
Sandra Schweizer,
Lincoln G. Scott,
J. Ho,
Scott Elrod,
Peter G. Schultz,
Richard A. Lerner,
Richard H. Bruce
Publication year - 2004
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.0403573101
Subject(s) - cytidine , isothermal microcalorimetry , biotin , enthalpy , chemistry , hexokinase , ligand (biochemistry) , streptavidin , biochemistry , biophysics , enzyme , biology , thermodynamics , glycolysis , physics , receptor
We report the fabrication of enthalpy arrays and their use to detect molecular interactions, including protein-ligand binding, enzymatic turnover, and mitochondrial respiration. Enthalpy arrays provide a universal assay methodology with no need for specific assay development such as fluorescent labeling or immobilization of reagents, which can adversely affect the interaction. Microscale technology enables the fabrication of 96-detector enthalpy arrays on large substrates. The reduction in scale results in large decreases in both the sample quantity and the measurement time compared with conventional microcalorimetry. We demonstrate the utility of the enthalpy arrays by showing measurements for two protein-ligand binding interactions (RNase A + cytidine 2'-monophosphate and streptavidin + biotin), phosphorylation of glucose by hexokinase, and respiration of mitochondria in the presence of 2,4-dinitrophenol uncoupler.