Intracellular Localization of Ca-binding protein MCBP-450 and the Regulatory Mechanism for Contraction in a Molluscan Smooth Muscle
Ca 结合蛋白 MCBP-450 的细胞内定位及软体动物平滑肌收缩的调节机制
基本信息
- 批准号:12640654
- 负责人:
- 金额:$ 2.37万
- 依托单位:
- 依托单位国家:日本
- 项目类别:Grant-in-Aid for Scientific Research (C)
- 财政年份:2000
- 资助国家:日本
- 起止时间:2000 至 2002
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
To make clear the intracellular localization of a Ca-binding protein, MCBP-450, and its regulatory mechanism for contraction, it has been thought that an experiment by immuno-electron microscopy is valid and significant. For the first step, in the present study, the isolation and purification of MCBP-450 from the anterior byssal retractor muscle (ABRM) of Mytilus edulis were examined to obtain the antigen prior to producing antibody for MCBP-450. Based on the method applied by the first report finding this protein (Yamanobe and Sugi, Biochim. Biophys. Acta 1149:166-174, 1993), a sample containing protein of molecular weight corresponding to 450 kDa was separated by SDS-PAGE. Ca-indicator quin2 was applied to the sample (Tatsumi et al., Ana. Biochem. 254:126-131, 1997). Then it was proved that the protein had an ability to bind Ca ions, by detecting the emission of fluorescence. It is reasonable to conclude that the sample is containing the MCBP-450. However, the protein yielded was ver … More y small in quantity, and difficult to supply directly for antibody producing. Therefore, the author tried to examine the amino acid sequence of this protein, in order to synthesize some peptides in vitro as an antigen for immunity. SDS-PAGE after the dialysis of purified samples showed no distinct band corresponding to 450 kDa, but a remarkable band corresponding to 〜100 kDa. The amino acid sequence of three peptides fragmented enzymatically from the 〜100 kDa protein was examined with Procise 494HT Protein Sequencing System, and determind the sequences of 12 amino acids, respectively. The homology-search of these sequences to the various known protein sequences using the NCBInr Library indicated a high homology (〜90%) with α-actinin of various kinds of cells and tissues. It is suggested that a part of MCBP-450 molecule resembles closely to some parts of α-scrinin. As a preliminary experiment, intracellular localization of a noble Ca-binding protein, calsequestrin, in ABRM fibers was also examined immuno-electron microscopically, and proved the localization of calsequestrin in the lumen of sarcoplasmic reticumum. Further experiments to reveal the intracellular localization of MCBP-450 is now in progress. Less
为了弄清钙结合蛋白MCBP-450在细胞内的定位及其收缩调节机制,免疫电子显微镜的实验被认为是有效的和有意义的。第一步,在本研究中,从紫贻贝前牵引肌(ABRM)中分离纯化MCBP-450,以获得MCBP-450的抗体。根据首次报道的方法,发现了该蛋白(Yamanobe和Sugi,Biochim)。生物群落。(Acta 1149:166-174,1993),用SDS-PAGE分离出相对分子质量为450 kDa的蛋白质。将钙指示剂Quin2应用于样品(TatSumi等人,Ana.生物化学。254:126-131,1997)。通过荧光检测,证明该蛋白具有结合钙离子的能力。可以合理地得出结论,样品中含有MCBP-450。然而,所产生的蛋白质是Ver…数量较少,难以直接供应抗体生产。因此,作者试图对该蛋白的氨基酸序列进行分析,以便在体外合成一些多肽作为免疫抗原。纯化样品透析后的SDS-PAGE显示在450 kDa处没有明显的条带,而在~100 kDa处有一条明显的条带。用Procise 494HT蛋白质测序系统测定了从~100 kDa蛋白质中酶切下来的3个多肽的氨基酸序列,并分别测定了12个氨基酸的序列。利用NCBInr文库对这些序列进行同源性搜索,发现它们与各种细胞和组织的α-肌动蛋白有很高的同源性(~90%)。提示MCBP-450分子的一部分与α-丝蛋白的某些部分非常相似。作为初步实验,我们还用免疫电子显微镜观察了钙结合蛋白在ABRM纤维中的细胞内定位,证实了钙结合蛋白在肌浆网管腔中的定位。进一步的实验揭示了MCBP-450的细胞内定位,目前正在进行中。较少
项目成果
期刊论文数量(21)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
鈴木季直: "生物学データ大百科事典(下巻、石原勝敏他編)"朝倉書店. 1462-1503 (2002)
铃木时直:“生物数据百科全书(第2卷,石原胜俊等编)”朝仓书店1462-1503(2002)。
- DOI:
- 发表时间:
- 期刊:
- 影响因子:0
- 作者:
- 通讯作者:
Suzuki,Suechika.: "Ultrasturucrural analysis of fiber formation during shoot development in Kenaf plants. I. Electron microscope studies on the cell wall formation in the cell of shoot apex (in Japanese)"Iann. Rep. Res. Ins. For Integrated Sci. Kanagawa U
Suzuki,Suechika.:“红麻植物芽发育过程中纤维形成的超声波分析。I.芽尖细胞细胞壁形成的电子显微镜研究(日语)”Iann。
- DOI:
- 发表时间:
- 期刊:
- 影响因子:0
- 作者:
- 通讯作者:
Suzuki, Suechika: "Ultrastructural organization of the transverse tubules and the sarcoplasmic reticulum in a fish sound-producing muscle"J. Electron Microsc.. 52(in press). (2003)
Suzuki,Suechika:“鱼发声肌肉中横管和肌浆网的超微结构组织”J。
- DOI:
- 发表时间:
- 期刊:
- 影响因子:0
- 作者:
- 通讯作者:
鈴木季直: "ハンドブック、生物学データ集(中村裕昭他編)"朝倉書店(印刷中). (2001)
铃木时直:“手册,生物数据收集(中村宏明等人编辑)”朝仓书店(目前正在印刷)(2001年)。
- DOI:
- 发表时间:
- 期刊:
- 影响因子:0
- 作者:
- 通讯作者:
Sugi,Haro.: "Possible contribution in hoseshoe crab skeletal muscle fibers"Adv. Exp. Med. Biol.. 481. 371-382 (2000)
Sugi,Haro.:“对软管蟹骨骼肌纤维的可能贡献”Adv.
- 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 }}
SUZUKI Suechika其他文献
SUZUKI Suechika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('SUZUKI Suechika', 18)}}的其他基金
Cryosection-Elemental Analysis Studies on the Regulatory Mechanism of Contraction by Calcium bound to Inner Surface of Smooth Muscle Plasma Membrane
平滑肌质膜内表面钙结合的收缩调节机制的冰冻切片元素分析研究
- 批准号:
07670063 - 财政年份:1995
- 资助金额:
$ 2.37万 - 项目类别:
Grant-in-Aid for Scientific Research (C)