Comparative analysis of 10 small molecules binding to carbonic anhydrase II by different investigators using Biacore technology.
In this benchmark study, 26 investigators were asked to characterize the kinetics and affinities of 10 sulfonamide inhibitors binding to the enzyme carbonic anhydrase II using Biacore optical biosensors. A majority of the participants collected data that could be fit to a 1:1 interaction model, but a subset of the data sets obtained from some instruments were of poor quality. The experimental errors in the k(a), k(d), and K(D) parameters determined for each of the compounds averaged 34, 24, and 37%, respectively. As expected, the greatest variation in the reported constants was observed for compounds with exceptionally weak affinity and/or fast association rates. The binding constants determined using the biosensor correlated well with solution-based titration calorimetry measurements. The results of this study provide insight into the challenges, as well as the level of experimental variation, that one would expect to observe when using Biacore technology for small molecule analyses.
Papalia
Giuseppe A
GA
Center for Biomolecular Interaction Analysis, School of Medicine, University of Utah, Salt Lake City, UT 84132, USA.
Leavitt
Stephanie
S
Bynum
Maggie A
MA
Katsamba
Phinikoula S
PS
Wilton
Rosemarie
R
Qiu
Huawei
H
Steukers
Mieke
M
Wang
Siming
S
Bindu
Lakshman
L
Phogat
Sanjay
S
Giannetti
Anthony M
AM
Ryan
Thomas E
TE
Pudlak
Victoria A
VA
Matusiewicz
Katarzyna
K
Michelson
Klaus M
KM
Nowakowski
Agnes
A
Pham-Baginski
Anh
A
Brooks
Jonathan
J
Tieman
Bryan C
BC
Bruce
Barry D
BD
Vaughn
Michael
M
Baksh
Michael
M
Cho
Yun Hee
YH
Wit
Mieke De
MD
Smets
Alexandra
A
Vandersmissen
Johan
J
Michiels
Lieve
L
Myszka
David G
DG
D015374:Biosensing Techniques; D002151:Calorimetry; D024402:Carbonic Anhydrase II / Q000737:chemistry* / Q000378:metabolism*; D002257:Carbonic Anhydrase Inhibitors / Q000145:classification / Q000378:metabolism*; D015588:Observer Variation; D011485:Protein Binding; D012108:Research Personnel; D013449:Sulfonamides / Q000037:antagonists & inhibitors* / Q000145:classification; D020349:Surface Plasmon Resonance / Q000295:instrumentation / Q000592:standards
D002257:Carbonic Anhydrase Inhibitors; D013449:Sulfonamides; D024402:Carbonic Anhydrase II