摘要
借助蛋白质内源荧光和活力测定,考察pH、温度、NaF和二价金属阳离子等因素对溶液中磷酸酯酶构象的影响。其中在pH6.0以下,蛋白质的三级结构变得松散,荧光强度下降,pH5.0时尤为显著,以后随pH值升高其三级结构得到恢复。在10～30℃范围内,蛋白荧光峰位置和强度变化不大,而在40～90℃范围内荧光强度升高再下降。NaF强烈抑制此酶,不同浓度的NaF使蛋白质荧光发射光谱发生明显变化。二价阳离子调控此酶活性,但可能与荧光光谱所反映的三级结构变化无关。
The influence of pH, temperature, NaF and bivalent cations on the conformation of phosphatase in solution, was investigated by measuring the intrinsic fluorescence of phosphatase and the enzyme activity.It was founded that the tertiary structure loose and the intensity of fluorescence decreased below pH 6.0. The intensity of fluorescence is lowest at pH 5.0 and the tertiary structure is reconstructed as the solution pH is increased.The characters of fluorescence spectra is not changed in the temperature region of 10～30?℃. The fluorescence intensity of the phosphatase increased, followed by a decreasing process, in the temperature region of 40～90?℃. The enzyme is inhibited by NaF significantly and the fluorescence spectra of the protein is influenced obviously by the concentration of NaF. The result showes that the bivalent cations can modulate the activity of the phosphatase but the mechanism maybe is independent of the tertiary structure.
出处
《四川大学学报（工程科学版）》
EI
CAS
CSCD
2003年第3期74-78,共5页
Journal of Sichuan University (Engineering Science Edition)
基金
国家自然科学基金资助项目(39970068)
关键词
类囊体
磷酸酯酶
去磷酸化
荧光光谱
thylakoid
phosphatase
dephosphorylation
fluorescence emission spectrum
