Synthesis of Ca^<2+> -responsive DNA-binding Allosteric Protein
Ca^2-响应性DNA结合变构蛋白的合成
基本信息
- 批准号:03453117
- 负责人:
- 金额:$ 4.1万
- 依托单位:
- 依托单位国家:日本
- 项目类别:Grant-in-Aid for General Scientific Research (B)
- 财政年份:1991
- 资助国家:日本
- 起止时间:1991 至 1992
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
DNA-binding proteins are classified into a few structural motifs. One of them is an assembly of two chains with many basic residues, which are brought close with each other by the extended peptide fragment called "Leu zipper". In the present study, we have used cyclic peptides which were expected to mimic function of "Leu zipper". Two chains of dodecapeptides were connected to cyclic octapeptides, cyclo(Leu-Sar-Lys(Z)-Sar)_2 (C8KS) and cyclo(Leu-Sar-Lys(CHO)-Sar-Leu-Sar-Glu(OBzl)-Sar) (C8KE). Both cyclic peptides formed a complex with Ca^<2+>. The protecting groups of the cyclic peptides were removed, and two chains of Boc-Glu-Napala-Leu-Aib-(Lys-Aib-Leu-Aib)_2 - (Napala represents 2-naphthylalanine) were bound to C8KS (F12-C8KS), and Ac-Glu(OMe)-Trp-Leu-Aib-(Lys-Aib-Leu-Aib)_2-and-(Leu-Aib-Glu(ONe)-Aib)_2-Lew-Aib-Ant (Ant represents an anthryl derivative) to C8KE (CH2). CD and fluorescence spectroscopy revealed that CH2 took a super-secondary structure of association of two helical chains in a buffer solution. The structure should be stable due to the following two reasons: i) two chains protrude over the same side of the cyclic skeleton, and ii) the dodecapeptides take an amphiphilic alpha helical conformation.With the addition of Lambda DNA to CH2 in alpha buffer solution, the fluorescence from the Trp residue was quenched, and a new signal from the anthryl group appeared at a longer wavelength. Thus, CH2 interacted with Lambda DNA, probably intercalating the indolyl and anthryl groups into base pairs of the DNA. On the other hand, F12-C8KS and a cyclic peptide having one chain did not interact with DNA. Therefore, the super-secondary structure of CH2 should be critical for the interaction with DNA. Interestingly, the interaction mode of CH2 and DNA was changed by the presence of Ca^<2+>.
DNA结合蛋白分为几种结构基序。其中之一是由两条具有许多碱性残基的链组装而成的,这两条链通过称为“Leu拉链”的延伸肽片段彼此靠近。在本研究中,我们使用了预期模拟“Leu拉链”功能的环肽。将两条十二肽链连接到环状八肽环上,分别命名为cyclo(Leu-Sar-Lys(Z)-Sar)_2(C8 KS)和cyclo(Leu-Sar-Lys(CHO)-Sar-Leu-Sar-Glu(OBzl)-Sar)(C8 KE)。两种环肽都能与Ca^2+形成复合物。除去环肽的保护基,Boc-Glu-Napala-Leu-Aib-(Lys-Aib-Leu-Aib)_2 -(Napala代表2-萘基丙氨酸)的两条链与C8 KS结合(F12-C8 KS),Ac-Glu(OMe)-Trp-Leu-Aib-(Lys-Aib-Leu-Aib)_2-和-(Leu-Aib-Glu(ONe)-Aib)_2-Lew-Aib-Ant(Ant代表蒽基衍生物)的两条链与C8 KE(CH 2)结合。CD和荧光光谱表明,在缓冲溶液中,CH 2采取了超二级结构的两个螺旋链的关联。由于以下两个原因,该结构应该是稳定的:i)两个链突出于环状骨架的同一侧,ii)十二肽采取两亲性α螺旋构象。在α缓冲溶液中,将λ DNA加入到CH 2中,来自Trp残基的荧光被猝灭,并且在较长波长处出现来自蒽基的新信号。因此,CH 2与λ DNA相互作用,可能将吲哚基和蒽基插入DNA的碱基对中。另一方面,F12-C8 KS和具有一条链的环肽不与DNA相互作用。因此,CH 2的超二级结构对于与DNA的相互作用是至关重要的。有趣的是,Ca^<2+>的存在改变了CH 2与DNA的相互作用模式。
项目成果
期刊论文数量(2)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Yukio Imanishi and Shunsaku Kimura(分担執筆): "Fundamental Investigations on the Creation of Biofunctional" 化学同人, 9 (1991)
今西幸雄和木村俊作(合著者):“生物功能创造的基础研究” Kagaku Doujin,9(1991)
- DOI:
- 发表时间:
- 期刊:
- 影响因子:0
- 作者:
- 通讯作者:
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}
数据更新时间:{{ journalArticles.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ monograph.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ sciAawards.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ conferencePapers.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ patent.updateTime }}
IMANISHI Yukio其他文献
IMANISHI Yukio的其他文献
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}
{{ truncateString('IMANISHI Yukio', 18)}}的其他基金
APPLICATION OF BIOMATERIALS IMMOBILIZED WITH BIOSIGNAL MOLECULES
生物信号分子固定化生物材料的应用
- 批准号:
07505023 - 财政年份:1995
- 资助金额:
$ 4.1万 - 项目类别:
Grant-in-Aid for Scientific Research (A)
JAPAN-ITALY PROGRAM ON FRONTIER FIELDS -NATURE AND HUMAN LIFE-
日本-意大利前沿领域-自然与人类生活-计划
- 批准号:
05045029 - 财政年份:1993
- 资助金额:
$ 4.1万 - 项目类别:
Grant-in-Aid for international Scientific Research
Japan-Italy Program on Frontier Fields Nature and Human Life
日本-意大利自然与人类生活前沿领域项目
- 批准号:
03045028 - 财政年份:1991
- 资助金额:
$ 4.1万 - 项目类别:
Grant-in-Aid for international Scientific Research
Photoresponsive Membrane-Associated Mutant Enzyme Prepared by Semisynthesis
半合成光响应膜相关突变酶
- 批准号:
01470112 - 财政年份:1989
- 资助金额:
$ 4.1万 - 项目类别:
Grant-in-Aid for General Scientific Research (B)