Chemical-genetic inhibition of sensitized mutant unconventional myosins.

Chemical-genetic inhibition of sensitized mutant unconventional myosins.
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致敏突变型非常规肌球蛋白的化学遗传抑制。

DOI:
10.1007/978-1-59745-490-2_16
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发表时间:
2007
期刊:
Methods in molecular biology (Clifton, N.J.)
影响因子:
--
通讯作者:
Mercer,JohnA
Mercer,JohnA
中科院分区:
--
文献类型:
--
作者:
Karcher,RyanL;Provance,DWilliam;Gillespie,PeterG;Mercer,JohnA

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敏化突变体非常规肌球蛋白的构建是一种在具有部分重叠功能的相关肌球蛋白背景下确定单个肌球蛋白功能的极好方法。在本章中,我们概述了通过突变使肌球蛋白敏感并针对核苷酸类似物组进行筛选的步骤,包括转染、显微注射和体外肌动蛋白共沉淀。我们还描述了在突变体和同源类似物被识别后设计功能实验所涉及的条件和考虑因素。当化学遗传学的强大策略正确应用于非常规肌球蛋白时,可以通过出色的内部控制对目标运动进行特异性和选择性抑制。
The construction of a sensitized mutant unconventional myosin is an excellent method for determining the function of the individual myosin against a background of related myosins with partially overlapping functions. In this chapter, we outline the steps involved in sensitizing myosin by mutation and screening them against panels of nucleotide analogs, including transfection, microinjection, and actin co-sedimentation in vitro. We also describe conditions and considerations involved in designing functional experiments after a mutant and cognate analog have been identified. The powerful strategy of chemical genetics, when correctly applied to unconventional myosins, enables both the specific and selectable inhibition of the target motor with outstanding internal controls.
DOI: 10.1073/pnas.93.20.10826
发表时间: 1996-10-01
影响因子: 11.1
作者:
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通讯作者: Mercer, JA
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DOI: --
发表时间: 1985
期刊: Journal of applied biochemistry
影响因子: --
作者:
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