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钙/钙调蛋白依赖性蛋白激酶IIβ亚型在轴突生长过程中于运动神经元中表达,并且是慢速轴突运输的一部分。

Calcium/calmodulin-dependent protein kinase IIbeta isoform is expressed in motor neurons during axon outgrowth and is part of slow axonal transport.

作者信息

Lund Linda M, McQuarrie Irvine G

机构信息

VA Medical Center, Cleveland, Ohio, USA.

出版信息

J Neurosci Res. 2002 Mar 15;67(6):720-8. doi: 10.1002/jnr.10162.

Abstract

Previously, we identified calcium/calmodulin-dependent protein kinase IIbeta (CaMKIIbeta) mRNA in spinal motor neurons with 372 bp inserted in what corresponds to the "association" domain of the protein. This was interesting because known additions and deletions to CaMKIIbeta mRNA are usually less than 100 bp in size and found in the "variable" region. Changes in the association domain of CaMKIIbeta could influence substrate specificity, activity or intracellular targeting. We show that three variations of this insert are found in CNS neurons or sciatic motor neurons of Sprague-Dawley rats. We used PCR and nucleic acid sequencing to identify inserts of 114, 243, or 372 bases. We also show that addition of the 372 bases is associated with outgrowth of the axon (the standard CaMKIIbeta downregulates when axon outgrowth occurs). Radiolabeling, immunoblots, and 2D PAGE identified this larger CaMKIIbeta as part of the group of soluble proteins moving at the slowest rate of axonal transport (SCa) in sciatic motor neurons (similar1 mm/day). This group is composed mainly of structural proteins (e.g., tubulin) used to assemble the cytoskeleton of regrowing axons.

摘要

此前,我们在脊髓运动神经元中鉴定出钙/钙调蛋白依赖性蛋白激酶IIβ(CaMKIIβ)信使核糖核酸,其在与该蛋白“结合”结构域相对应的位置插入了372个碱基对。这很有意思,因为已知CaMKIIβ信使核糖核酸的增减通常大小小于100个碱基对,且出现在“可变”区域。CaMKIIβ结合结构域的变化可能会影响底物特异性、活性或细胞内靶向作用。我们发现,在斯普拉格-道利大鼠的中枢神经系统神经元或坐骨神经运动神经元中存在这种插入片段的三种变体。我们使用聚合酶链反应(PCR)和核酸测序来鉴定114、243或372个碱基的插入片段。我们还发现,添加372个碱基与轴突生长有关(当轴突生长发生时,标准的CaMKIIβ会下调)。放射性标记、免疫印迹和二维聚丙烯酰胺凝胶电泳(2D PAGE)确定这种更大的CaMKIIβ是坐骨神经运动神经元中轴突运输速率最慢的可溶性蛋白组(SCa)的一部分(速度约为1毫米/天)。该蛋白组主要由用于组装再生轴突细胞骨架的结构蛋白(如微管蛋白)组成。

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