Dynamic motions of ice-binding proteins in living Caenorhabditis elegans using diffracted X-ray blinking and tracking.
Dynamic motions of ice-binding proteins in living Caenorhabditis elegans using diffracted X-ray blinking and tracking.
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DOI：
10.1016/j.bbrep.2022.101224
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发表时间：
2022-03
影响因子：
2.7
通讯作者：
Sasaki YC
中科院分区：
文献类型：
--
作者：
Kuramochi M;Dong Y;Yang Y;Arai T;Okada R;Shinkai Y;Doi M;Aoyama K;Sekiguchi H;Mio K;Tsuda S;Sasaki YC
蛋白质分子的动力学性质涉及其结构与功能之间的关系。时间分辨的X射线观测能够以皮米级的精度捕获生物分子的结构。然而，这项技术还没有在活体动物中实施。在这里，我们研究了衍射X射线闪烁（DXB）和衍射X射线跟踪（DXT），以观察位于成年秀丽隐杆线虫肠细胞上的蛋白质的动态。在20 °C至−10 °C的温度下，对这种体内组织特异性DXB进行了检测，以获得通过配备有甘氨酸-丝氨酸接头的跨膜CD 4蛋白与细胞连接的来自舌形番茄嗜冷菌（AnpIBP）的重组冰结合蛋白。AnpIBP通过与冰晶结合来抑制冰在零度以下的生长。我们发现AnpIBP的旋转运动在−10 °C时减小。相比之下，具有冰结合能力缺陷的AnpIBP突变体的运动在-10 °C时没有减少。在5 °C以上，DXT法测得AnpIBP的扭转和倾斜运动速度均高于缺陷型AnpIBP。这些结果表明，野生型AnpIBP是高度移动的在溶液中，它是通过冰结合在零度以下的温度停止。DXB和DXT允许以亚纳米分辨率精度探索活体动物中的蛋白质行为。本文用活体DXB和DXT技术分析了冰结合蛋白（AnpIBP）的动态运动。AnpIBP的旋转运动在-10 °C时减少，而缺陷型AnpIBP突变体在-10 °C时没有减少。在高于5 °C的温度下测量的AnpIBP的扭转和倾斜运动总是高于缺陷型AnpIBP突变体的扭转和倾斜运动。
The dynamic properties of protein molecules are involved in the relationship between their structure and function. Time-resolved X-ray observation enables capturing the structures of biomolecules with picometre-scale precision. However, this technique has yet to be implemented in living animals. Here, we examined diffracted X-ray blinking (DXB) and diffracted X-ray tracking (DXT) to observe the dynamics of a protein located on intestinal cells in adult Caenorhabditis elegans. This in vivo tissue-specific DXB was examined at temperatures from 20 °C to −10 °C for a recombinant ice-binding protein from Antarctomyces psychrotrophicus (AnpIBP) connected with the cells through a transmembrane CD4 protein equipped with a glycine-serine linker. AnpIBP inhibits ice growth at subzero temperatures by binding to ice crystals. We found that the rotational motion of AnpIBP decreases at −10 °C. In contrast, the motion of the AnpIBP mutant, which has a defective ice-binding ability, did not decrease at −10 °C. The twisting and tilting motional speeds of AnpIBPs measured above 5 °C by DXT were always higher than those of the defective AnpIBP mutant. These results suggest that wild-type AnpIBP is highly mobile in solution, and it is halted at subzero temperatures through ice binding. DXB and DXT allow for exploring protein behaviour in live animals with subnano resolution precision. Dynamic motions of ice-binding protein (AnpIBP) were analysed in live animals using in vivo DXB and DXT techniques. The rotational motion of AnpIBP decreases at −10 °C, whereas that of the defective AnpIBP mutant did not decrease at −10 °C. The twisting and tilting motions of AnpIBPs measured above 5 °C were always higher than those of the defective AnpIBP mutant.
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影响因子：
4.6
作者：
Sekiguchi H;Suzuki Y;Nishino Y;Kobayashi S;Shimoyama Y;Cai W;Nagata K;Okada M;Ichiyanagi K;Ohta N;Yagi N;Miyazawa A;Kubo T;Sasaki YC
通讯作者：
Sasaki YC
影响因子：
3.8
作者：
Chang, Jaewon;Nishijima, Masaki;Sasaki, Yuji C.
通讯作者：
Sasaki, Yuji C.
影响因子：
3.4
作者：
Jang TH;Park SC;Yang JH;Kim JY;Seok JH;Park US;Choi CW;Lee SR;Han J
通讯作者：
Han J
影响因子：
56.9
作者：
CHALFIE, M;TU, Y;PRASHER, DC
通讯作者：
PRASHER, DC
影响因子：
3.7
作者：
Lee H
通讯作者：
Lee H