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dc.contributor.authorRivas-Pardo, Jaime Andrés
dc.contributor.authorLi, Yong
dc.contributor.authorMártonfalvi, Zsolt
dc.contributor.authorTapia-Rojo, Rafael
dc.contributor.authorUnger, Andreas
dc.contributor.authorFernández-Trasancos, Ángel
dc.contributor.authorHerrero-Galan, Elas 
dc.contributor.authorVelazquez-Carreras, Diana 
dc.contributor.authorFernández, Julio M
dc.contributor.authorLinke, Wolfgang A
dc.contributor.authorAlegre-Cebollada, Jorge
dc.identifier.citationNat Commun. 2020; 11(1):2060es_ES
dc.description.abstractSingle-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.es_ES
dc.description.sponsorshipThis work was supported by the National Institutes of Health grants GM116122 and HL61228 (J.M.F.). J.A.R.P. acknowledges funding from CONICYT grant 11180705. J. A.C. acknowledges funding from the Ministerio de Ciencia e Innovación through grants BIO2014-54768-P, BIO2017-83640-P, and RYC-2014-16604, and the Regional Government of Madrid (S2018/NMT-4443). The CNIC is supported by the Instituto de Salud Carlos III (ISCIII), the Ministerio de Ciencia e Innovación and the Pro CNIC Foundation, and is a Severo Ochoa Center of Excellence (SEV-2015-0505). W.A.L. acknowledges funding from the German Research Foundation (SFB1002, TPA08) and IZKF Munster (Li1/029/20). J.A.C. and W.A.L. acknowledge funding from the European Research Area Network on Cardiovascular Disease through consortium MINOTAUR (ISCIII-AC16/00045). Z.M. acknowledges funding from the National Research, Development and Innovation Office (NKFIH) through grants PD116558, FK128956, NVKP-16-1-2016-0017, the Hungarian Academy of Sciences Bolyai grant and an EMBO short term fellowship (ASTF 360-2016). A.F.T. was the recipient of a travel fellowship from the Boehringer Ingelheim Fonds.es_ES
dc.publisherSpringer es_ES
dc.titleA HaloTag-TEV genetic cassette for mechanical phenotyping of proteins from tissues.es_ES
dc.typejournal articlees_ES
dc.rights.licenseAtribución 4.0 Internacional*
dc.contributor.funderMinisterio de Ciencia e Innovación (España) 
dc.contributor.funderComunidad de Madrid (España) 
dc.contributor.funderInstituto de Salud Carlos III 
dc.contributor.funderFundación ProCNIC 
dc.contributor.funderDeutsche Forschungsgemeinschaft (Alemania) 
dc.contributor.funderHungarian Academy of Sciences 
dc.contributor.funderEuropean Molecular Biology Organization 
dc.contributor.funderBoehringer Ingelheim Fonds 
dc.identifier.journalNature communicationses_ES
dc.repisalud.orgCNICCNIC::Grupos de investigación::Mecánica molecular del sistema cardiovasculares_ES
dc.rights.accessRightsopen accesses_ES

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