Interactions of microtubule-associated protein MAP2 with unpolymerized and polymerized tubulin and actin using a 96-well microtiter plate solid-phase immunoassay.
Interactions of microtubule-associated protein MAP2 with unpolymerized and polymerized tubulin and actin using a 96-well microtiter plate solid-phase immunoassay.
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使用 96 孔微量滴定板固相免疫测定法观察微管相关蛋白 MAP2 与未聚合和聚合的微管蛋白和肌动蛋白的相互作用。
作者：
B. Pedrotti;R. Colombo;K. Islam
固相免疫分析法用于研究微管相关蛋白MAP 2与细胞骨架蛋白微管蛋白和肌动蛋白之间的蛋白质-蛋白质相互作用。该测定可在96孔微量滴定板上进行，并可用于研究与亚基蛋白及其各自的聚合物、微管和微丝的相互作用。微量滴定格式允许处理大量样品，并且可以改变许多条件。在该固相免疫测定中，MAP 2以浓度依赖性方式与微管/微丝和微管蛋白二聚体/G-肌动蛋白结合。然而，即使在高NaCl浓度下，结合的MAP 2也不从丝上解离，而同时加入NaCl减少了MAP 2与这些蛋白质的结合。MgCl 2是1个数量级更有效地降低MAP 2结合相比，NaCl，这表明MAP 2可能通过“屏蔽”微管蛋白二聚体之间的静电排斥。还检查了MAP 2在交联微丝和微管中的作用。微管/微管蛋白结合的MAP 2表现出降低的能力，结合到微丝和G-肌动蛋白，而微丝/G-肌动蛋白结合的MAP 2能够有效地结合到微管和微管蛋白二聚体。（250字处删节）
A solid-phase immunoassay is used to study the protein-protein interactions between microtubule-associated protein MAP2 and the cytoskeletal proteins tubulin and actin. The assay can be performed on 96-well microtiter plates and can be used to study the interactions with both subunit proteins and their respective polymers, microtubules and microfilaments. The microtiter format allows a large number of samples to be processed, and a number of conditions can be varied. In this solid-phase immunoassay MAP2 bound to microtubules/microfilaments and tubulin dimers/G-actin in a concentration-dependent manner. However, the bound MAP2 was not dissociated from the filaments even at high NaCl concentrations, while simultaneous addition of NaCl diminished MAP2 binding to these proteins. MgCl2 was 1 order of magnitude more efficient in decreasing MAP2 binding compared with NaCl, suggesting that MAP2 may act by "screening" the electrostatic repulsion between tubulin dimers. The role of MAP2 in cross-linking microfilaments and microtubules was also examined. Microtubule/tubulin-bound MAP2 showed a diminished ability to bind to both microfilaments and G-actin, while microfilament/G-actin-bound MAP2 was able to bind efficiently to both microtubules and tubulin dimers.(ABSTRACT TRUNCATED AT 250 WORDS)
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微管相关蛋白2与微管的平衡结合机制。
DOI：
--
发表时间：
1993
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Wallis,KT;Azhar,S;Rho,MB;Lewis,SA;Cowan,NJ;Murphy,DB
通讯作者：
Murphy,DB
通过自缔合和肌动蛋白结合大分子交联肌动蛋白丝网络。
DOI：
--
发表时间：
1982
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Griffith,LM;Pollard,TD
通讯作者：
Pollard,TD
无需组装促进剂即可对微管进行可逆组装纯化，并进一步纯化微管蛋白、微管相关蛋白和 MAP 片段。
影响因子：
--
作者：
Vallee,RB
通讯作者：
Vallee,RB
微管相关蛋白 2 的广泛 cAMP 依赖性和 cAMP 非依赖性磷酸化。
DOI：
--
发表时间：
1983
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Theurkauf,WE;Vallee,RB
通讯作者：
Vallee,RB
DOI：
10.1016/0006-291x(80)91175-4
发表时间：
1980-01-01
影响因子：
3.1
作者：
MACLEANFLETCHER, S;POLLARD, TD
通讯作者：
POLLARD, TD