Dynamic motions of ice-binding proteins in living Caenorhabditis elegans using diffracted X-ray blinking and tracking

• Published in: Biochemistry and Biophysics Reports
• Main Authors: Masahiro Kuramochi, Yige Dong, Yue Yang, Tatsuya Arai, Rio Okada, Yoichi Shinkai, Motomichi Doi, Kouki Aoyama, Hiroshi Sekiguchi, Kazuhiro Mio, Sakae Tsuda, Yuji C. Sasaki
• Format: Article in Journal/Newspaper
• Language: English
• Published: Elsevier 2022
• Subjects: Diffracted X-ray tracking (DXT); Diffracted X-ray blinking (DXB); C. elegans; Ice-binding protein; Biology (General); QH301-705.5; Biochemistry; QD415-436; Tilting; Antarc*
• Online Access: https://doi.org/10.1016/j.bbrep.2022.101224; https://doaj.org/article/d51b7b0d33ea4dc7a01a263f0631b0b5

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.