蜡样芽孢杆菌磷脂酶C的构象研究。尿素对该酶的影响。
Conformational studies on phospholipase C from Bacillus cereus. The effect of urea on the enzyme.
作者信息
Little C
出版信息
Biochem J. 1978 Dec 1;175(3):977-86. doi: 10.1042/bj1750977.
Abstract
- When heated in 8 M-urea, phospholipase C(EC 3.1.4.3) from Bacillus cereus undergoes conformational transitions depending on the temperatures used. These transitions were studied by examining protein fluorescence, iodide quenching of protein fluorescence, u.v. difference spectroscopy, chemical availability of histidine residues in the enzyme, circular dichroism and catalytic activity. 2. Unless simultaneously exposed to elevated temperatures the enzyme appears to be unaffected by 8 M-urea. Removal of the two zinc atoms from the enzyme renders phospholipase C very sensitive to denaturation by 8 M-urea as indicated by fluorescence emission spectra and circular dichroism. 3. Both the native and the zinc-free enzymes are markedly more resistant to irreversible thermal inactivation in the presence of 8 M-urea than in its absence. 4. The response of the enzyme to 8 M-urea and the role of zinc in stabilizing the enzyme are discussed.
摘要
- 蜡样芽孢杆菌的磷脂酶C（EC 3.1.4.3）在8M尿素中加热时，会根据所用温度发生构象转变。通过检测蛋白质荧光、蛋白质荧光的碘化物猝灭、紫外差光谱、酶中组氨酸残基的化学可及性、圆二色性和催化活性来研究这些转变。2. 除非同时暴露于高温下，该酶似乎不受8M尿素的影响。从酶中去除两个锌原子后，如荧光发射光谱和圆二色性所示，磷脂酶C对8M尿素引起的变性非常敏感。3. 无论是天然酶还是无锌酶，在存在8M尿素的情况下都比不存在时对不可逆热失活具有明显更高的抗性。4. 讨论了该酶对8M尿素的反应以及锌在稳定酶方面的作用。
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