2026-06-14T03:29:09Zhttps://repisalud.isciii.es/rest/oai/requestoai:repisalud.isciii.es:20.500.12105/103212024-10-31T11:45:13Zcom_20.500.12105_19604com_20.500.12105_2051col_20.500.12105_19605
Repisalud
author
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
funder
Ministerio de Ciencia e Innovación (España)
funder
Comunidad de Madrid (España)
funder
Instituto de Salud Carlos III
funder
Fundación ProCNIC
funder
Deutsche Forschungsgemeinschaft (Alemania)
funder
Hungarian Academy of Sciences
funder
European Molecular Biology Organization
funder
Boehringer Ingelheim Fonds
2020-06-10T09:54:47Z
2020-06-10T09:54:47Z
2020-04-28
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
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.
eng
A HaloTag-TEV genetic cassette for mechanical phenotyping of proteins from tissues.
journal article
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