Kinetics of head-tail joining in bacteriophage T4D studied by quasi-elastic light scattering: effects of temperature, pH, and ionic strength.

Kinetics of head-tail joining in bacteriophage T4D studied by quasi-elastic light scattering: effects of temperature, pH, and ionic strength.
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通过准弹性光散射研究噬菌体 T4D 中头尾连接的动力学:温度、pH 和离子强度的影响。

DOI:
10.1021/bi00520a031
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发表时间:
1981
期刊:
影响因子:
2.9
通讯作者:
Bloomfield,VA
Bloomfield,VA
中科院分区:
生物学3区
文献类型:
--
作者:
Aksiyote-Benbasat,J;Bloomfield,VA

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唐纳德岛Stimpson、Julyet Aksiyote-Benbasat和维克托A.布卢姆菲尔德摘要:我们已经确定了温度,pH值和离子强度对纯化的T4 D头部和尾部的附着动力学的影响,目的是了解这种病毒组装反应的一些机械细节。为了产生用于反应的头部,我们使用缺乏尾部和尾部纤维的抗渗透压休克突变体(10”/18“/19”/34”/36 '/37“);对于尾部,使用13”/23“突变体。我们推测,基因13的突变是防止连接器连接到头部和尾部,从而阻止它们连接所必需的。为了跟踪反应,我们使用准弹性光散射(QLS)[Aksiyote-Benbasat,J.,&布卢姆菲尔德,弗吉尼亚州(1975)分子生物学杂志(J. Mol. Biol. 95,335-357],实际上测量了当尾部连接时头部的平均平移扩散系数的降低。头部和尾部浓度分别在6 X 1010和1.2 X 1011个颗粒/mL范围内,相当于约(1或2)X 10 - 10 M。由于磁头的高散射能力,这种极端的稀释是可能的,它将非常快速的双分子反应减慢到^ 2= 400 s。我们能够每50秒测量一次QLS自相关函数。
Donald I. Stimpson, Julyet Aksiyote-Benbasat, and Victor A. Bloomfield abstract: We have determined the effects of temperature, pH, and ionic strength on the kinetics of attachment of purified T4D heads and tails, with the aim of understanding some of the mechanistic details of this viral assembly reaction. To produce heads for the reaction, we used an osmotic shock resistant mutant deficient in tails and tail fibers (10"/18 “/19"/34"/36'/37 “); for tails, a 13"/23 “mutant was used. We conjecture that the mutation in gene 13 was necessary to prevent connectors from being attached to both heads and tails, thus blocking theirjoining. To follow the reaction, we used quasi-elastic light scattering (QLS)[Aksiyote-Benbasat, J., & Bloomfield, VA (1975) J. Mol. Biol. 95, 335-357], in effect measuring the decrease in average translational diffusion coefficient of theheads as tails are attached. The head and tail concentrations were in the range 6 X 1010 and 1.2 X 1011 particles/mL, respectively, corresponding to about (1 or 2) X 10~ 10 M. This extreme dilution, which is possible because of the high scattering power of the heads, slows down the very rapid bimolecular reaction to^ 2= 400 s. We were able to measure the QLS autocorrelation function once every 50 s.
噬菌体 T4 头部次要蛋白质成分的定位
DOI: 10.1128/jvi.24.1.121-134.1977
发表时间: 1977
影响因子: 5.4
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