Nanoimages show disruption of tubulin polymerization by chlorpyrifos oxon: implications for neurotoxicity.
Nanoimages show disruption of tubulin polymerization by chlorpyrifos oxon: implications for neurotoxicity.
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DOI：
10.1016/j.taap.2009.07.015
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发表时间：
2009-10-15
影响因子：
3.8
通讯作者：
Lockridge O
中科院分区：
文献类型：
--
作者：
Grigoryan H;Lockridge O
有机磷制剂会导致某些人的认知缺陷和抑郁症。我们推测，有机磷制剂引起这些疾病的机制是通过改变大脑中的蛋白质。一种这样的蛋白质可以是微管蛋白。微管蛋白聚合形成微管，将细胞成分运送到神经轴突。本工作的目的是测量有机磷试剂毒死蜱对微管蛋白聚合的影响。另一个目标是确定由毒死蜱oxon在微管聚合物中共价修饰的氨基酸，并将其与未聚合的微管蛋白二聚体中修饰的氨基酸进行比较。在室温下用0.005 - 0.1mM毒死蜱氧磷处理纯化的牛微管蛋白（0.1mM）30分钟，然后通过加入1mM GTP聚合以产生微管。通过原子力显微镜观察微管。采用串联离子阱电喷雾电离和基质辅助激光解吸/电离质谱技术对毒死蜱氧磷修饰的胰蛋白酶肽进行了鉴定。纳米成像显示，低浓度（0.005和0.01毫米）的毒死蜱oxon产生短，薄的微管。0.025 mM的浓度刺激聚合，而高浓度（0.05和0.1 mM）引起聚集。在未聚合的微管蛋白二聚体中的17个酪氨酸被毒死蜱oxon共价修饰，在聚合的微管中只有2个酪氨酸被标记。聚合的微管蛋白中的两个标记的酪氨酸是α微管蛋白的EDAANNY * R中的Tyr 103和β微管蛋白的GSQQY * R中的Tyr 281。总之，毒死蜱氧磷结合微管蛋白破坏微管蛋白聚合。这些结果可能导致有机磷制剂的神经毒性的理解。
Organophosphorus agents cause cognitive deficits and depression in some people. We hypothesize that the mechanism by which organophosphorus agents cause these disorders is by modification of proteins in the brain. One such protein could be tubulin. Tubulin polymerizes to make the microtubules that transport cell components to nerve axons. The goal of the present work was to measure the effect of the organophosphorus agent chlorpyrifos oxon on tubulin polymerization. An additional goal was to identify the amino acids covalently modified by chlorpyrifos oxon in microtubule polymers and to compare them to the amino acids modified in unpolymerized tubulin dimers. Purified bovine tubulin (0.1 mM) was treated with 0.005-0.1 mM chlorpyrifos oxon for 30 min at room temperature and then polymerized by addition of 1 mM GTP to generate microtubules. Microtubules were visualized by atomic force microscopy. Chlorpyrifos oxon-modified residues were identified by tandem ion trap electrospray ionization and matrix-assisted laser desorption/ionization mass spectrometry of tryptic peptides. Nanoimaging showed that low concentrations (0.005 and 0.01 mM) of chlorpyrifos oxon yielded short, thin microtubules. A concentration of 0.025 mM stimulated polymerization, while high concentrations (0.05 and 0.1 mM) caused aggregation. Of the 17 tyrosines covalently modified by chlorpyrifos oxon in unpolymerized tubulin dimers, only 2 tyrosines were labeled in polymerized microtubules. The two labeled tyrosines in polymerized tubulin were Tyr 103 in EDAANNY*R of alpha tubulin, and Tyr 281 in GSQQY*R of beta tubulin. In conclusion, chlorpyrifos oxon binding to tubulin disrupts tubulin polymerization. These results may lead to an understanding of the neurotoxicity of organophosphorus agents.
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DOI：
10.1097/01.jom.0000235938.70212.dd
发表时间：
2006-10-01
影响因子：
3.2
作者：
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通讯作者：
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影响因子：
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作者：
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影响因子：
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作者：
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影响因子：
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作者：
Hancock, Dana B.;Martin, Eden R.;Mayhew, Gregory M.;Stajich, Jeffrey M.;Jewett, Rita;Stacy, Mark A.;Scott, Burton L.;Vance, Jeffery M.;Scott, William K.
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影响因子：
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