FEBS Letters | |
Pyrene excimer fluorescence as a proximity probe for investigation of residual structure in the unfolded state of human carbonic anhydrase II | |
Hammarström, Per1  Carlsson, Uno1  Jonsson, Bengt-Harald2  Kalman, Björn1  | |
[1] IFM-Department of Chemistry, Linköping University, S-581 83 Linköping, Sweden;Department of Biochemistry, Umeå University, S-901 87 Umeå, Sweden | |
关键词: Excimer fluorescence; Pyrene; Folding; Carbonic anhydrase; Residual structure; DMSO; dimethylsulfoxide; GuHCl; guanidine hydrochloride; HCAII; human carbonic anhydrase II; PMIA; N-(1-pyrenemethyl)iodoacetamide; py-N67C/C206-py; pyrene-labeled cysteines in a HCAII mutant with a Asn-67→Cys-67 substitution; | |
DOI : 10.1016/S0014-5793(97)01488-9 | |
学科分类:生物化学/生物物理 | |
来源: John Wiley & Sons Ltd. | |
【 摘 要 】
The excimer fluorescence from two pyrenyl moieties attached to cysteines in human carbonic anhydrase II has been monitored to characterize residual structure retained under strong denaturing conditions. A position in β-strand 3, N67C, together with the single naturally occurring cysteine 206 in β-strand 7, were used as attachment sites. The eximer formation by the pyrenyls, requiring proximity of the probes, revealed an unfolding transition at a GuHCl concentration significantly higher than that required to induce unfolding of the molten globule state as monitored by CD. These results indicate that the excimer transition monitors the unfolding of a residual compact structure that spans β-strands 3–7. This region constitutes the central and the most hydrophobic part of the molecule, emphasizing the importance of hydrophobic interaction in maintaining residual structure under strong unfolding conditions.
【 授权许可】
Unknown
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