PMID- 24375583
OWN - NLM
STAT- MEDLINE
DCOM- 20140902
LR  - 20131230
IS  - 1099-1352 (Electronic)
IS  - 0952-3499 (Linking)
VI  - 27
IP  - 1
DP  - 2014 Jan
TI  - Label-free characterization of carbonic anhydrase-novel inhibitor interactions 
      using surface plasmon resonance, isothermal titration calorimetry and 
      fluorescence-based thermal shift assays.
PG  - 46-56
LID - 10.1002/jmr.2330 [doi]
AB  - This work describes the development of biophysical unbiased methods to study the 
      interactions between new designed compounds and carbonic anhydrase II (CAII) 
      enzyme. These methods have to permit both a screening of a series of sulfonamide 
      derivatives and the identification of a lead compound after a thorough study of 
      the most promising molecules. Interactions data were collected using surface 
      plasmon resonance (SPR) and thermal shift assay (TSA). In the first step, 
      experiments were performed with bovine CAII isoform and were extended to human 
      CAII. Isothermal titration calorimetry (ITC) experiments were also conducted to 
      obtain thermodynamics parameters necessary for the processing of the TSA data. 
      Results obtained with this reference methodology demonstrate the effectiveness of 
      SPR and TSA. KD values obtained from SPR data were in perfect accordance with 
      ITC. For TSA, despite the fact that the absolute values of KD were quite 
      different, the same affinity scale was obtained for all compounds. The binding 
      affinities of the analytes studied vary by more than 50 orders of magnitude; for 
      example, the KD value determined by SPR were 6 +/- 4 and 299 +/- 25 nM for compounds 
      1 and 3, respectively. This paper discusses some of the theoretical and 
      experimental aspects of the affinity-based methods and evaluates the protein 
      consumption to develop methods for the screening of further new compounds. The 
      double interest of SPR, that is, for screening and for the quick thorough study 
      of the interactions parameters (ka , kd , and KD ), leads us to choose this 
      methodology for the study of new potential inhibitors.
CI  - Copyright (c) 2013 John Wiley & Sons, Ltd.
FAU - Rogez-Florent, Tiphaine
AU  - Rogez-Florent T
AD  - Universite Lille Nord de France, F-59000, Lille, France; UDSL, EA-4481, UFR 
      Pharmacie, Universite Lille Nord de France, F-59000, Lille, France.
FAU - Duhamel, Laetitia
AU  - Duhamel L
FAU - Goossens, Laurence
AU  - Goossens L
FAU - Six, Perrine
AU  - Six P
FAU - Drucbert, Anne-Sophie
AU  - Drucbert AS
FAU - Depreux, Patrick
AU  - Depreux P
FAU - Danze, Pierre-Marie
AU  - Danze PM
FAU - Landy, David
AU  - Landy D
FAU - Goossens, Jean-Francois
AU  - Goossens JF
FAU - Foulon, Catherine
AU  - Foulon C
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - J Mol Recognit
JT  - Journal of molecular recognition : JMR
JID - 9004580
RN  - 0 (Carbonic Anhydrase Inhibitors)
RN  - EC 4.2.1.- (Carbonic Anhydrase II)
SB  - IM
MH  - Animals
MH  - *Calorimetry
MH  - Carbonic Anhydrase II/*metabolism
MH  - Carbonic Anhydrase Inhibitors/*pharmacology
MH  - Cattle
MH  - *Fluorescence
MH  - Humans
MH  - Kinetics
MH  - Protein Binding
MH  - *Surface Plasmon Resonance
MH  - *Thermodynamics
OTO - NOTNLM
OT  - binding
OT  - carbonic anhydrase
OT  - diarylpyrazole sulfonamide
OT  - fluorescence-based thermal shift assay
OT  - isothermal titration calorimetry
OT  - kinetics
OT  - surface plasmon resonance
OT  - thermodynamics
EDAT- 2014/01/01 06:00
MHDA- 2014/09/03 06:00
CRDT- 2013/12/31 06:00
PHST- 2013/05/28 00:00 [received]
PHST- 2013/09/19 00:00 [revised]
PHST- 2013/09/28 00:00 [accepted]
PHST- 2013/12/31 06:00 [entrez]
PHST- 2014/01/01 06:00 [pubmed]
PHST- 2014/09/03 06:00 [medline]
AID - 10.1002/jmr.2330 [doi]
PST - ppublish
SO  - J Mol Recognit. 2014 Jan;27(1):46-56. doi: 10.1002/jmr.2330.