title: Enthalpy arrays.
abstract: 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.
authors.affiliation: Scripps-PARC Institute for Advanced Biomedical Sciences, Palo Alto Research Center, 3333 Coyote Hill Road, Palo Alto, CA 94304, USA.
authors.forename: Francisco E
authors.initials: FE
authors.lastname: Torres
authors.forename: Peter
authors.initials: P
authors.lastname: Kuhn
authors.forename: Dirk
authors.initials: D
authors.lastname: De Bruyker
authors.forename: Alan G
authors.initials: AG
authors.lastname: Bell
authors.forename: Michal V
authors.initials: MV
authors.lastname: Wolkin
authors.forename: Eric
authors.initials: E
authors.lastname: Peeters
authors.forename: James R
authors.initials: JR
authors.lastname: Williamson
authors.forename: Gregory B
authors.initials: GB
authors.lastname: Anderson
authors.forename: Gregory P
authors.initials: GP
authors.lastname: Schmitz
authors.forename: Michael I
authors.initials: MI
authors.lastname: Recht
authors.forename: Sandra
authors.initials: S
authors.lastname: Schweizer
authors.forename: Lincoln G
authors.initials: LG
authors.lastname: Scott
authors.forename: Jackson H
authors.initials: JH
authors.lastname: Ho
authors.forename: Scott A
authors.initials: SA
authors.lastname: Elrod
authors.forename: Peter G
authors.initials: PG
authors.lastname: Schultz
authors.forename: Richard A
authors.initials: RA
authors.lastname: Lerner
authors.forename: Richard H
authors.initials: RH
authors.lastname: Bruce
mesh_terms: D019297:2,4-Dinitrophenol / Q000494:pharmacology; D000818:Animals; D001710:Biotin / Q000737:chemistry / Q000378:metabolism*; D002417:Cattle; D019069:Cell Respiration / Q000187:drug effects; D003568:Cytidine Monophosphate / Q000737:chemistry / Q000378:metabolism*; D004867:Equipment Design; D005947:Glucose / Q000378:metabolism; D006593:Hexokinase / Q000378:metabolism; D008024:Ligands; D008929:Mitochondria, Heart / Q000187:drug effects / Q000378:metabolism; D010766:Phosphorylation; D040081:Protein Array Analysis / Q000295:instrumentation / Q000379:methods*; D011485:Protein Binding; D012259:Ribonuclease, Pancreatic / Q000737:chemistry / Q000378:metabolism*; D019809:Streptavidin / Q000737:chemistry / Q000378:metabolism*; D013816:Thermodynamics
chemical_list: D008024:Ligands; D001710:Biotin; D019809:Streptavidin; D006593:Hexokinase; D012259:Ribonuclease, Pancreatic; D003568:Cytidine Monophosphate; D005947:Glucose; D019297:2,4-Dinitrophenol