Abstract
The effects of guanidine hydrochloride and high temperature on human glycophorin and sialic acid-free glycophorin were monitored by circular dichroism, viscosity, and fluorescence of 1-anilino-8-naphthalane sulfonate (ANS). The following observations were made: 1. 1.|Glycophorin and its sialic acid-free counterpart are unusually stable to both guanidine · HCl and heat. 2. 2.|CD and viscosity measurements indicate that guanidine · HCl neither causes a cooperative unfolding nor generates a random coil. 3. 3.|The ANS binding site is much more sensitive to guanidine · HCl than the ellipticity at 220 nm ( θ 220). 4. 4.|The effect of temperature on CD is reversible whereas the effect of guanidine · HCl is not. 5. 5.|The carbohydrate moiety influences the viscosity, and also contributes to the changes in θ 220 when solutions of glycophorin are heated. These unusual properties indicate a complex mechanism of unfolding for this structurally stable macromolecule.
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