Folding of egg white proteins as a post-translational processing.
蛋清蛋白的折叠作为翻译后加工。
基本信息
- 批准号:63560086
- 负责人:
- 金额:$ 1.22万
- 依托单位:
- 依托单位国家:日本
- 项目类别:Grant-in-Aid for General Scientific Research (C)
- 财政年份:1988
- 资助国家:日本
- 起止时间:1988 至 1989
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
Egg white proteins, ovotransferrin and ovalbumin, which are synthesized in hen oviducts under hormonal regulation, were investigated for their folding mechanisms using in vitro translation system as well as refolding systems of denatured forms.A two-step procedure was found to be useful for the efficient refolding of a complex protein, ovotransferrin. In the first step, the reduced and denatured form of the protein was incubated at a low temperature in a nondenaturing buffer containing reduced glutathione; in the second step, the reduced form was reoxidized at a higher temperature in the presence of oxidized glutathione. Under these conditions, the fully reduced forms of ovotransferrin and its half-molecules were almost quantitatively reoxidized to regain iron-binding abilities and conformations, very similar to the native form. The circular dichroism spectra revealed that at low temperatures the fully reduced forms have partially folded conformations, which are fluctuating like "molten globule" states. The reoxidization kinetics compared between whole ovotransferrin and the two half-molecules supported independent refolding of the N- and C-terminal domains.With respect to ovalbumin about 40% of urea-denatured protein was renatured to the native form after 18 hr incubation under non-denaturing conditions. Likewise, only a part of the mRNA-directed translation product in wheat germ translation system was found to take a native-like conformation.
利用体外翻译系统和变性复性系统研究了荷尔蒙调节下母鸡输卵管合成的卵清蛋白、卵转铁蛋白和卵清蛋白的折叠机制。在第一步中,将还原和变性的蛋白质在含有还原型谷胱甘肽的非变性缓冲液中低温孵育;在第二步中,在氧化型谷胱甘肽的存在下,还原形式在更高的温度下重新氧化。在这种条件下,完全还原的转铁蛋白及其半分子几乎被定量地重新氧化,以恢复与天然形式非常相似的铁结合能力和构象。圆二色谱表明,在低温下,完全还原的形式具有部分折叠的构象,这些构象像熔化的球状状态一样波动。比较全卵转铁蛋白和两个半分子之间的氧化动力学,支持N-末端和C-末端结构域的独立复性。对于卵清蛋白,在非变性条件下孵育18小时后,约40%的尿素变性蛋白复性为天然形式。同样,在小麦胚芽翻译系统中,只有一部分由mRNA引导的翻译产物具有天然的构象。
项目成果
期刊论文数量(7)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
H.Oe,N.Takahashi,E.Doi,& M.Hirose: "Effects of anion binding on the conformations of the two domains of ovotransferrin" Journal of Biochemistry. 106. 858-863 (1989)
H.Oe、N.Takahashi、E.Doi、
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M. Hirose, T. Akuta & N. Takahashi: "Renaturation of ovotransferrin under two-step conditions allowing primary folding of the fully reduced form and the subsequent regeneration of the intramolecular disulfides." The Journal of Biological Chemistry 264, 16
M.广濑,T.阿库塔
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H. Oe, N. Takahashi, E. Doi & M. Hirose: "Effects of anion binding on the conformations of the domains of ovotransferrin." Journal of Biochemistry 106, 858-863 (1989).
H. Oe、N. Takahashi、E. Doi
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M.Hirose,T.Akuta,& N.Takahashi: "Renaturation of ovotransferrin under two-step conditions allowing primary folding of fully reduced from and the subsequent regeration of the intramolecular disulfides" The Journal of Biological Chemistry. 264. 16867-16872
M.广濑,T.阿库塔,
- DOI:
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- 影响因子:0
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M.Hirose,T.Akuta,& N.Takahashi: "Renaturation of ovotransferrin under two-step conditions allowing primary folding of the fully reduced form and the subsequent regeneration of the intramolecular disulfides." The Journal of Biological Chemistry. 264. 16867
M.广濑,T.阿库塔,
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HIROSE Masaaki其他文献
HIROSE Masaaki的其他文献
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{{ truncateString('HIROSE Masaaki', 18)}}的其他基金
Protein engineering for conferring a biological function on ovalbumin
赋予卵清蛋白生物学功能的蛋白质工程
- 批准号:
15380229 - 财政年份:2003
- 资助金额:
$ 1.22万 - 项目类别:
Grant-in-Aid for Scientific Research (B)
Structural basis for the functional properties of food Proteins
食品蛋白质功能特性的结构基础
- 批准号:
10460057 - 财政年份:1998
- 资助金额:
$ 1.22万 - 项目类别:
Grant-in-Aid for Scientific Research (B)
Functional properties and conformational changes of food proteins - Roles of molten globule state
食品蛋白质的功能特性和构象变化 - 熔球状态的作用
- 批准号:
05453170 - 财政年份:1993
- 资助金额:
$ 1.22万 - 项目类别:
Grant-in-Aid for General Scientific Research (B)
Involvement of molten globule state on the folding process of secretary proteins
熔球状态对分泌蛋白折叠过程的影响
- 批准号:
02660094 - 财政年份:1990
- 资助金额:
$ 1.22万 - 项目类别:
Grant-in-Aid for General Scientific Research (C)
Studies of the hormone-dependent expression of conalbumin gene
伴清蛋白基因激素依赖性表达的研究
- 批准号:
61560096 - 财政年份:1986
- 资助金额:
$ 1.22万 - 项目类别:
Grant-in-Aid for General Scientific Research (C)
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