Effects of cryoprotectants on the structure and thermostability of the human carbonic anhydrase II-acetazolamide complex
| Autor: | Bhargav Kondeti, Chingkuang Tu, Mayank Aggarwal, David N. Silverman, Christopher D. Boone, Robert McKenna |
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| Rok vydání: | 2012 |
| Předmět: |
Glycerol
Models Molecular Sucrose Cryoprotectant Protein Conformation Carbonic anhydrase II Kinetics Crystallography X-Ray Carbonic Anhydrase II chemistry.chemical_compound Cryoprotective Agents Structural Biology Carbonic anhydrase Catalytic Domain Enzyme Stability medicine Humans Carbonic Anhydrase Inhibitors Thermostability biology Calorimetry Differential Scanning Temperature Active site General Medicine Research Papers Acetazolamide Crystallography chemistry biology.protein Biophysics medicine.drug |
| Zdroj: | Acta crystallographica. Section D, Biological crystallography. 69(Pt 5) |
| ISSN: | 1399-0047 |
| Popis: | Protein X-ray crystallography has seen a progressive shift from data collection at cool/room temperature (277–298 K) to data collection at cryotemperature (100 K) because of its ease of crystal preparation and the lessening of the detrimental effects of radiation-induced crystal damage, with 20–25%(v/v) glycerol (GOL) being the preferred choice of cryoprotectant. Here, a case study of the effects of cryoprotectants on the kinetics of carbonic anhydrase II (CA II) and its inhibition by the clinically used inhibitor acetazolamide (AZM) is presented. Comparative studies of crystal structure, kinetics, inhibition and thermostability were performed on CA II and its complex with AZM in the presence of either GOL or sucrose. These results suggest that even though the cryoprotectant GOL was previously shown to be directly bound in the active site and to interact with AZM, it affects neither the thermostability of CA II nor the binding of AZM in the crystal structure or in solution. However, addition of GOL does affect the kinetics of CA II, presumably as it displaces the water proton-transfer network in the active site. |
| Databáze: | OpenAIRE |
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