Thermodynamic benchmark study using Biacore technology.
A total of 22 individuals participated in this benchmark study to characterize the thermodynamics of small-molecule inhibitor-enzyme interactions using Biacore instruments. Participants were provided with reagents (the enzyme carbonic anhydrase II, which was immobilized onto the sensor surface, and four sulfonamide-based inhibitors) and were instructed to collect response data from 6 to 36 degrees C. van't Hoff enthalpies and entropies were calculated from the temperature dependence of the binding constants. The equilibrium dissociation and thermodynamic constants determined from the Biacore analysis matched the values determined using isothermal titration calorimetry. These results demonstrate that immobilization of the enzyme onto the sensor surface did not alter the thermodynamics of these interactions. This benchmark study also provides insights into the opportunities and challenges in carrying out thermodynamic studies using optical biosensors.
Navratilova
Iva
I
Center for Biomolecular Interaction Analysis, School of Medicine, University of Utah, Salt Lake City, UT 84132, USA.
Papalia
Giuseppe A
GA
Rich
Rebecca L
RL
Bedinger
Daniel
D
Brophy
Susan
S
Condon
Brad
B
Deng
Ta
T
Emerick
Anne W
AW
Guan
Hann-Wen
HW
Hayden
Tanya
T
Heutmekers
Thomas
T
Hoorelbeke
Bart
B
McCroskey
Mark C
MC
Murphy
Mary M
MM
Nakagawa
Terry
T
Parmeggiani
Fabio
F
Qin
Xiaochun
X
Rebe
Sabina
S
Tomasevic
Nenad
N
Tsang
Tiffany
T
Waddell
M Brett
MB
Zhang
Fred Feiyu
FF
Leavitt
Stephanie
S
Myszka
David G
DG
D019985:Benchmarking; D035843:Biomedical Research; D015374:Biosensing Techniques / Q000295:instrumentation* / Q000592:standards; D002151:Calorimetry / Q000295:instrumentation* / Q000592:standards*; D024402:Carbonic Anhydrase II / Q000737:chemistry / Q000378:metabolism; D002257:Carbonic Anhydrase Inhibitors / Q000145:classification* / Q000378:metabolism*; D015588:Observer Variation; D011485:Protein Binding; D013449:Sulfonamides / Q000037:antagonists & inhibitors* / Q000145:classification; D020349:Surface Plasmon Resonance / Q000295:instrumentation / Q000592:standards; D013816:Thermodynamics
D002257:Carbonic Anhydrase Inhibitors; D013449:Sulfonamides; D024402:Carbonic Anhydrase II