Molecular Assembly and the role in infection of Tail-lysozyme from bacteriophage T4.

分子组装及其在噬菌体 T4 尾部溶菌酶感染中的作用。

基本信息

  • 批准号:
    07680712
  • 负责人:
  • 金额:
    $ 1.41万
  • 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:
    1995
  • 资助国家:
    日本
  • 起止时间:
    1995 至 1996
  • 项目状态:
    已结题

项目摘要

Tail-lysozyme, gp5^<**>, is a 42 kD structural protein of the baseplate which has a lysozyme activity and is located under the baseplate. Upon adsorption of the phage to Escherichia coli, it penetrates into the outer membrane together with the tail tube and digs a whole on the peptidoglycan layr so that the tail tube can reach the inner membrane. We have previously isolated the tail-lysozyme and shown that the tail-lysozyme has the same substrate specificity as that of T4 lysozyme, gp e, namely N-acetyl muramidase (1). The expected molecular weight from the nucleotide sequence was 62 kD and in fact it is expressed as a 62 kD protein from the cloned gene 5, but the lysozyme activity is absent in the precurser protein (2). In the present study, we have determined the N-and C-terminal sequence of the mature protein. The N-terminal sequence coincided with that expected from the nucleotide sequence and the the C-terminus was Val390. A region with high homology (44% identical) to gp e, is located towards the C-terminus of the mature gp 5. N-terminal region is likely to assume a domain which constitutes the integral part of the baseplate. From the result, it was concluded that the 20 kD fragment is cleaved off from the precurser protein to become mature 42 kD tail-lysozyme.On the other hand, we have mapped the mutation sites of 12 mutants which have a mutation in gene 5. These include five ts (temperature sensitive), two hs (heat sensitive) and five amber mutants. All the mutation sites were mapped upstream of Val390. Among the ts mutants, 5ts1 (3) is a bypass e mutant wich does not require gp e to lyse from within. This was Gly322Asp mapped in the lysozyme domain. Another ts mutant, 5DH6318, is a bypass 63 mutant which does not require gp63 for efficient attachment of the tail fibers. This was Ala65Thr mapped in the "structural" domain. Including these mutations, other mutations will be discussed in respect to the structure-function relation of the tail-lysozyme.
Tail-lysozyme,gp5^&lt;**&gt;是一种位于基板下方的具有溶菌酶活性的42kD结构蛋白。噬菌体吸附到大肠杆菌后,与尾管一起穿透到外膜,并在肽聚糖上挖出一个整体,使尾管能够到达内膜。我们以前已经分离到了尾部溶菌酶,并证明了尾部溶菌酶与T4溶菌酶具有相同的底物特异性,即N-乙酰溶菌酰胺酶(1)。核苷酸序列的预期分子量为62kD,实际上表达为克隆基因5的62kD蛋白,但前递归蛋白(2)中没有溶菌酶活性。在本研究中,我们测定了成熟蛋白的N-端和C-端序列。N端序列与预期核苷酸序列一致,C端为Val390。与Gp e同源性高(44%相同)的区域位于成熟Gp 5的C-末端,N-末端区域可能承担构成基板组成部分的结构域。另一方面,我们对5号基因突变的12个突变株进行了突变定位,其中包括5个温度敏感突变株、2个hs突变株和5个琥珀突变株。所有突变位点均位于Val390的上游。在ts突变体中,5ts1(3)是一个旁路e突变体,不需要gp e从内部裂解。Gly322Asp定位于溶菌酶结构域。另一个ts突变体5DH6318是一个旁路63突变体,它不需要gp63来有效地附着尾部纤维。这是Ala65Thr被映射到“结构”域中。包括这些突变在内,还将讨论尾部溶菌酶的结构-功能关系方面的其他突变。

项目成果

期刊论文数量(23)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Takayo Sasaki, Tomoyuki Shinomiya, Takashi Kumazaki, Nobuko Mohri, Shin-ichi Ishii and Fumio Arisaka: "Nucleotide Sequences of the Contractile Tail Sheath and Tube Genes of Bacteriophage PS17 and Amino Acid Sequences of Their Products." Res.Comm.Biochem.&
Takayo Sasaki、Tomoyuki Shinomiya、Takashi Kumazaki、Nobuko Mohri、Shin-ichi Ishii 和 Fumio Arisaka:“噬菌体 PS17 的收缩尾鞘和管基因的核苷酸序列及其产品的氨基酸序列。”
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    0
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武田茂樹: "ネガティブ染色" 細胞工学. 16(6月号 印刷中). (1997)
Shigeki Takeda:“负染色”细胞工程 16(六月号印刷)(1997 年)。
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    0
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Hisayuki Morii: "Identification of Kinesin Neck Region as a Stable α-Helical Coiled-Coil and Its Thermodynamic Charcterization" Biochemistry. (in press). (1997)
Hisayuki Morii:“驱动蛋白颈部区域作为稳定 α-螺旋卷曲线圈的识别及其热力学表征”生物化学(1997 年)。
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    0
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  • 通讯作者:
Takuro Matsui, Bronya Riniviene, Edward Goldberg, Akira Tsugita, Nobuo Tanaka and Fumio Arisaka: "Isolation and Characterization of a Molecular Chaperone, gp57A,of Bacteriopahge T4" J.Bacteriol. 179. 1846-1851 (1997)
Takuro Matsui、Bronya Riniviene、Edward Goldberg、Akira Tsugita、Nobuo Tanaka 和 Fumio Arisaka:“Bacteriopahge T4 分子伴侣 gp57A 的分离和表征”J.Bacteriol。
  • DOI:
  • 发表时间:
  • 期刊:
  • 影响因子:
    0
  • 作者:
  • 通讯作者:
Hisayuki Morii, Tatsuyuki Takenawa, Fumio Arisaka, and Takashi Shimizu: "Identification of Kinesin Neck Region as a Stable alpha-Helical Coiled-Coil and Its Thermodynamic Charcterization." Biochemistry. 36. 1933-1942 (1997)
Hisayuki Morii、Tatsuyuki Takenawa、Fumio Arisaka 和 Takashi Shimizu:“驱动蛋白颈部区域作为稳定 α 螺旋卷曲线圈的识别及其热力学表征。”
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ARISAKA Fumio其他文献

ARISAKA Fumio的其他文献

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{{ truncateString('ARISAKA Fumio', 18)}}的其他基金

Structure and assembly of the central hub of the baseplate of bacteriophage T4
噬菌体T4基板中心轮毂的结构与组装
  • 批准号:
    23570190
  • 财政年份:
    2011
  • 资助金额:
    $ 1.41万
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
Structure Formation of the Neck which Links the Head and the Tail of Bacteriophage
连接噬菌体头部和尾部的颈部结构形成
  • 批准号:
    18570147
  • 财政年份:
    2006
  • 资助金额:
    $ 1.41万
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
Assembly and Mechanism of Infection of Bacteriophage
噬菌体的组装及感染机制
  • 批准号:
    16087204
  • 财政年份:
    2004
  • 资助金额:
    $ 1.41万
  • 项目类别:
    Grant-in-Aid for Scientific Research on Priority Areas
Structure and Assembly of the Contractile Tail of Bacteriophage
噬菌体收缩尾部的结构与组装
  • 批准号:
    15370065
  • 财政年份:
    2003
  • 资助金额:
    $ 1.41万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
Architecture and Principle of Formation of the Baseplate of Bacteriophage T4
噬菌体T4底板的结构和形成原理
  • 批准号:
    13680736
  • 财政年份:
    2001
  • 资助金额:
    $ 1.41万
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
X-ray Crystallographic Analysis of Bacteriophage Precursor Structures
噬菌体前体结构的 X 射线晶体学分析
  • 批准号:
    10044070
  • 财政年份:
    1998
  • 资助金额:
    $ 1.41万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B).
Architecture and Principle of Formation of the Baseplate of Bacteriophage T4
噬菌体T4底板的结构和形成原理
  • 批准号:
    10480178
  • 财政年份:
    1998
  • 资助金额:
    $ 1.41万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B).
Analyses of the Relationship between Amino Acid Substitution and Phenotype of the Tail Sheath Protein of Bacteriophage T4
噬菌体T4尾鞘蛋白氨基酸取代与表型关系分析
  • 批准号:
    02680125
  • 财政年份:
    1990
  • 资助金额:
    $ 1.41万
  • 项目类别:
    Grant-in-Aid for General Scientific Research (C)

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