Karabinos Anton, Schünemann Jürgen, Parry David A D
SEMBID, s.r.o.-Research Centre of Applied Biomedical Diagnostics, Magnezitarska 2/C, Kosice, 04013, Slovakia.
Department of Cellular Logistics, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, Goettingen, 37077, Germany.
Cytoskeleton (Hoboken). 2017 Mar;74(3):107-113. doi: 10.1002/cm.21354. Epub 2017 Jan 22.
The dimerisation properties of six intestine-expressed intermediate filament (IF) proteins (B2, C1, C2, D1, D2, E1) were analysed in blot overlay assay on membranes containing all of the eleven recombinant C. elegans IF proteins (A1, A2, A3, A4, B1, B2, C1, C2, D1, D2, and E1). The interactions detected in the blot assays exclusively comprise intestine-expressed IF proteins and the protein A4, which is found in the dauer larva intestine. About 86% of these interactions are heterotypic, while the remaining interactions relate to C1, C2, and D2 homodimers. These multiple modes of interaction were also supported by calculations of the numbers of possible interchain ionic interactions derived from the individual rod sequences. The results predict that the six B2, C1, C2, D1, D2, and E1 IF proteins are able to form as many as eleven different heteropolymeric and three homopolymeric IFs in the C. elegans intestine. This simple model of the intestinal IF meshwork enables us to speculate that our previously reported triple RNAi worms arrested or decreased their growth because of feeding reduction due to morphological defects of the mechanically compromised intestine.
在含有所有11种重组秀丽隐杆线虫中间丝（IF）蛋白（A1、A2、A3、A4、B1、B2、C1、C2、D1、D2和E1）的膜上进行印迹覆盖分析，以分析六种在肠道表达的中间丝（IF）蛋白（B2、C1、C2、D1、D2、E1）的二聚化特性。印迹分析中检测到的相互作用仅包括在肠道表达的IF蛋白和在 dauer 幼虫肠道中发现的蛋白A4。这些相互作用中约86%是异型的，而其余的相互作用与C1、C2和D2同二聚体有关。从各个杆状序列推导的可能链间离子相互作用数量的计算也支持了这些多种相互作用模式。结果预测，六种B2、C1、C2、D1、D2和E1 IF蛋白能够在秀丽隐杆线虫肠道中形成多达11种不同的异聚体IF和三种同聚体IF。这种简单的肠道IF网络模型使我们能够推测，我们之前报道的三重RNA干扰蠕虫由于机械受损肠道的形态缺陷导致摄食减少而停止生长或生长减缓。
