guanidine hydrochloride denaturation and refolding of human placental alkaline phosphatase（盐酸胍变性和重折叠的人类胎盘碱性磷酸酶）.docVIP

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guanidine hydrochloride denaturation and refolding of human placental alkaline phosphatase（盐酸胍变性和重折叠的人类胎盘碱性磷酸酶）.doc

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guanidine hydrochloride denaturation and refolding of human placental alkaline phosphatase（盐酸胍变性和重折叠的人类胎盘碱性磷酸酶）

Guanidine hydrochloride denaturation and refolding of human placental alkaline phosphatase Paper to write network: Author: Yanxia Geyin Lin Li Xin Geng Fang Song [Keywords:] hydrochloride [Abstract] Objective To observe the human placental alkaline phosphatase (HPAP in the guanidine hydrochloride denaturation and refolding process structure as dynamic changes in its folding mechanism. Methods spectrofluorimetric detection HPAP conformational changes detected by spectrophotometry of its vitality changes in the results of low concentrations of guanidine hydrochloride (lt;0.5 mol / L), the enzyme fluorescence intensity decreased slightly, the maximum emission wavelength unchanged at 340 nm, and enzyme activity was enhanced with the increase in the concentration of guanidine hydrochloride, the fluorescence intensity increased, the obvious red shift of the maximum emission wavelength, the rapid decline in enzyme activity, when the GuHCl concentration increased to 3.0 mol / L, the maximum emission wavelength to stop at 365 nm, the loss of enzyme activity, but the fluorescence intensity of the guanidine concentration to 5.0 mol / L before to stop the change. conclusion HPAP in the degeneration process of the tri-state, denatured intermediate state enzyme renaturation, the denatured enzyme is only part of the refolding, and refolding conditions. [Keywords:] alkaline phosphatase; guanidine hydrochloride; refolding; degeneration [ABSTRACT] ObjectiveTo examine the changes of conformation and activity of human placental alkaline phosphatase (HPAP) in denaturation and renaturation of guanidine chloride, and to study its folding mechanism.MethodsSpectrofluorimetry and spectrophotometry was used to determine the changes of conformation and activity of HPAP, respectively.ResultsThe fluorescence emission intensity of low concentration guanidine chloride (lower than 0.5 mol / L) decreased a little, without any red shift of emission maximum, and the wave length remained