Effects of cryoprotectants on the structure and thermostability of the human carbonic anhydrase II–acetazolamide complex
- University of Florida, 1600 SW Archer Road, PO Box 100245, Gainesville, FL 32610 (United States)
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. 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.
- OSTI ID:
- 22351286
- Journal Information:
- Acta Crystallographica. Section D: Biological Crystallography, Vol. 69, Issue Pt 5; Other Information: PMCID: PMC3640473; PMID: 23633596; PUBLISHER-ID: wd5202; OAI: oai:pubmedcentral.nih.gov:3640473; Copyright (c) International Union of Crystallography 2013; Country of input: International Atomic Energy Agency (IAEA); ISSN 0907-4449
- Country of Publication:
- Denmark
- Language:
- English
Similar Records
ENHANCEMENT OF THE RENAL EXCRETION OF CESIUM-137 IN RATS TREATED WITH ACETAZOLAMIDE AND RELATED COMPOUNDS
Carbonic anhydrase-inhibitor binding: From solution to the gas phase