Characterization of folding intermediates during urea-induced denaturation of human carbonic anhydrase II.

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Wahiduzzaman, Dar MA, Haque MA, Idrees D, Hassan MI, Islam A, Ahmad F

Characterization of folding intermediates during urea-induced denaturation of human carbonic anhydrase II.

Int J Biol Macromol. 2017 Feb;95:881-887. doi: 10.1016/j.ijbiomac.2016.10.073. Epub 2016 Oct 24.

PubMed ID

27789330 [ View in PubMed

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Abstract

Knowledge of folding/unfolding pathway is fundamental basis to study protein structure and stability. Human carbonic anhydrase II (HCAII) is a approximately 29kDa, beta-sheet dominated monomeric protein of 259 amino acid residues. In the present study, the urea-induced denaturation of HCAII was carried out which was a tri-phasic process, i.e., N (native) <--> XI <--> XII <--> D (denatured) with stable intermediates XI and XII populated around 2 and 4M urea, respectively. The far-UV CD was used to characterize the intermediate states (XI and XII) for secondary structural content, near-UV CD for tertiary structure, dynamic light scattering for hydrodynamic radius and ANS fluorescence spectroscopy for the presence of exposed hydrophobic patches. Based on these experiments, we concluded that urea-induced XI state has characteristics of molten globule state while XII state bears characteristics features of pre-molten globule state. Characterization of the intermediates on the folding pathway will contribute to a deeper understanding of the structure-function relationship of HCAII. Furthermore, this system may provide an excellent model to study urea stress and the strategies adopted by the organisms to combat such a stress.

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Drug Targets

Drug	Target	Kind	Organism	Pharmacological Action	Actions
Urea	Carbonic anhydrase 2	Protein	Humans	Unknown	Not Available	Details