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- W2000542088 endingPage "425" @default.
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- W2000542088 abstract "The effects of desiccation and rehydration cycles encountered by extracellular enzymes in soils are studied on α-chymotrypsin adsorbed on montmorillonite. The controlled hygrometric FTIR cell used in this study enables to monitor drying and rehydration steps undergone by the α-chymotrypsin-montmorillonite suspension or by the enzyme alone. Relative humidity (RH) determines the amount of deuterated water in the FTIR cell atmosphere. The molar water/protein ratio (W/P) as well as the conformational and solvation states of the enzyme have been determined using H/D exchange monitored by FTIR-transmission spectroscopy. When the W/P ratio decreases from 3500 to ∼400, unfolding of β-secondary structure in three different domains involves about 8% of the polypeptide backbone with respect to the most solvated states. Desiccation induces β-unfolding, which opens channels allowing free vapor water molecules to diffuse into the enzyme at 15% RH. On drying to 0% RH, displacements of internal water (H2O) in the enzyme are demonstrated by reverse peptide isotopic exchanges (COND ⇒ CONH). Specific β-structures, only formed in highly solvated states, sequester around 20 internal H2O molecules. Indeed, most of the unfolded secondary structures during the drying step are refolded at W/P∼1000 during rehydration. However, self-association hinders the recovery of the initial closed tertiary structure. The pD-dependent structural changes controlling inward and outward water diffusion are suppressed, whether the protein is initially in an adsorbed state or in solution. Changes in secondary structures encountered during desiccation/rehydration cycle are similar for the protein either free or in the adsorbed state. Thus domains that are unfolded by adsorption are not concerned by the desiccation/rehydration cycle." @default.
- W2000542088 created "2016-06-24" @default.
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- W2000542088 date "2004-05-01" @default.
- W2000542088 modified "2023-09-25" @default.
- W2000542088 title "Structural effects of drying and rehydration for enzymes in soils: a kinetics-FTIR analysis of α-chymotrypsin adsorbed on montmorillonite" @default.
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- W2000542088 doi "https://doi.org/10.1016/j.jcis.2004.01.067" @default.
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