2026-06-14T03:30:33Zhttps://repisalud.isciii.es/rest/oai/request

oai:repisalud.isciii.es:20.500.12105/103212024-10-31T11:45:13Zcom_20.500.12105_19604com_20.500.12105_2051col_20.500.12105_19605

00925njm 22002777a 4500 dc Rivas-Pardo, Jaime Andrés author Li, Yong author Mártonfalvi, Zsolt author Tapia-Rojo, Rafael author Unger, Andreas author Fernández-Trasancos, Ángel author Herrero-Galan, Elas author Velazquez-Carreras, Diana author Fernández, Julio M author Linke, Wolfgang A author Alegre-Cebollada, Jorge author 2020-04-28 Single-molecule methods using recombinant proteins have generated transformative hypotheses on how mechanical forces are generated and sensed in biological tissues. However, testing these mechanical hypotheses on proteins in their natural environment remains inaccesible to conventional tools. To address this limitation, here we demonstrate a mouse model carrying a HaloTag-TEV insertion in the protein titin, the main determinant of myocyte stiffness. Using our system, we specifically sever titin by digestion with TEV protease, and find that the response of muscle fibers to length changes requires mechanical transduction through titin's intact polypeptide chain. In addition, HaloTag-based covalent tethering enables examination of titin dynamics under force using magnetic tweezers. At pulling forces < 10 pN, titin domains are recruited to the unfolded state, and produce 41.5 zJ mechanical work during refolding. Insertion of the HaloTag-TEV cassette in mechanical proteins opens opportunities to explore the molecular basis of cellular force generation, mechanosensing and mechanotransduction. Nat Commun. 2020; 11(1):2060 10.1038/s41467-020-15465-9 2041-1723 Nature communications 32345978 http://hdl.handle.net/20.500.12105/10321 A HaloTag-TEV genetic cassette for mechanical phenotyping of proteins from tissues.