Structural properties of recombinant ovalbumin and its transformation into a thermostabilized form by alkaline treatment.
Structural properties of recombinant ovalbumin and its transformation into a thermostabilized form by alkaline treatment.
复制标题
重组卵清蛋白的结构特性及其通过碱处理转化为热稳定形式。
DOI：
10.1271/bbb.63.1392
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发表时间：
1999
期刊：
影响因子：
--
通讯作者：
M. Hirose
中科院分区：
文献类型：
--
作者：
Y. Arii;N. Takahashi;E. Tatsumi;M. Hirose
从大肠杆菌细胞质中分离纯化重组卵清蛋白，并对其化学性质和构象性质进行分析。重组卵清蛋白表现出与蛋清几乎完全相同的圆二色性和固有色氨酸荧光光谱。根据氨基酸分析的结果，与蛋清蛋白一样，重组蛋白中含有4个半胱氨酸巯基和1个半胱氨酸二硫基；二硫键通过肽图谱分析发现与天然胱氨酸Cys73-Cys120对应。根据凝胶电泳分析，二硫键的存在是由细胞质蛋白纯化过程中相应的半胱氨酸残基的特异性氧化引起的。与构象和肽结构的同一性不同，在重组蛋白中没有检测到已知的蛋白蛋白的翻译后修饰（n端乙酰化、磷酸化和糖基化）。将重组卵清蛋白转化为热稳定形式，类似于将蛋清蛋白转化为s -卵清蛋白；碱性处理使卵白蛋白的热稳定性温度提高了8.7℃。这些数据有力地表明，卵白蛋白的翻译后修饰与s -卵白蛋白的形成机制无关。
The recombinant ovalbumin produced in Escherichia coli was purified from the cytoplasmic fraction and analyzed for its chemical and conformational properties. The recombinant ovalbumin displayed almost exactly the same circular dichroism and intrinsic tryptophan fluorescence spectra as egg white ovalbumin. As in the egg white protein, four cysteine sulfhydryls and one cystine disulfide were contained in the recombinant protein, according to the results of amino acid analyses; the disulfide bond was found by a peptide mapping analysis to correspond to the native cystine, Cys73-Cys120. According to a gel electrophoresis analysis, the presence of the disulfide bond was accounted for by specific oxidation of the corresponding cysteine residues during purification of the cytoplasmic protein. Unlike the identity in the conformational and peptide structures, none of the post-translational modifications (N-terminal acetylation, phosphorylation, and glycosylation) that are known with egg white ovalbumin were detected in the recombinant protein. The recombinant ovalbumin was transformed into a thermostabilized form in a similar manner to the transformation of egg white protein into S-ovalbumin; alkaline treatment increased the temperature for thermostability by 8.7 degrees C. These data strongly suggest that the post-translational modifications of ovalbumin are not related to the formation mechanism for S-ovalbumin.
DOI：
10.1073/pnas.90.3.1038
发表时间：
1993-02-01
影响因子：
11.1
作者：
BARDWELL, JCA;LEE, JO;BECKWITH, J
通讯作者：
BECKWITH, J