In spite of extensive work on the functional sequelae of Hypertrophic CardioMyopathy (HCM)-associated mutations in sarcomeric proteins, the mechanisms by which the mutant proteins cause the disease have not been definitely identified. Here we use the single myofibril technique (Tesi et al., Biophys. J., 2002, 83, 2142-2151) to compare the kinetics of contraction and relaxation of myofibrils isolated from frozen left ventricular samples of one homozygous HCM patient carrying the cardiac Troponin T (cTnT) mutation K273N (underwent heart transplantation) to those from “control” hearts. Preparations, mounted in a force recording apparatus (15 °C), were maximally Ca2+-activated (pCa 4.5) and fully relaxed (pCa 9) by rapid (<10 ms) solution switching. The rate constant of active tension generation following maximal Ca2+ activation (kACT) was markedly faster in the myofibrils from the K273N patient (1.7- 2 s−1) compared to controls (0.7-1 s−1). Replacement of the mutant protein by exchange with wild-type recombinant human cTn reduced kACT of HCM myofibrils close to control values (1 s−1). Force relaxation kinetics upon Ca2+ removal were also faster in K273N myofibrils than in controls, evidence that the apparent rate with which cross-bridges leave the force generating states is accelerated in the HCM preparations. The results indicate that the HCM-associated cTnT mutation K273N alters apparent cross-bridge kinetics. This can lead to increased energy cost of tension generation that may be central to the HCM disease process. Supported by the 7th Framework Program of the European Union, “BIG-HEART” grant agreement 241577, & Telethon-Italy GGP07133.
The HCM-Associated Cardiac Troponin T Mutation K273N Accelerates Tension Generation and Relaxation in Human Cardiac Myofibrils / Claudia Ferrara; Nicoletta Piroddi; Beatrice Scellini; Cecilia Ferrantini; Sabine J. van Dijk; Cris dos Remedios; Chiara Tesi; Jolanda van der Velden; Corrado Poggesi. - In: BIOPHYSICAL JOURNAL. - ISSN 0006-3495. - STAMPA. - 100(3):(2011), pp. 115a-115a.
The HCM-Associated Cardiac Troponin T Mutation K273N Accelerates Tension Generation and Relaxation in Human Cardiac Myofibrils
FERRARA, CLAUDIA;PIRODDI, NICOLETTA;SCELLINI, BEATRICE;FERRANTINI, CECILIA;TESI, CHIARA;POGGESI, CORRADO
2011
Abstract
In spite of extensive work on the functional sequelae of Hypertrophic CardioMyopathy (HCM)-associated mutations in sarcomeric proteins, the mechanisms by which the mutant proteins cause the disease have not been definitely identified. Here we use the single myofibril technique (Tesi et al., Biophys. J., 2002, 83, 2142-2151) to compare the kinetics of contraction and relaxation of myofibrils isolated from frozen left ventricular samples of one homozygous HCM patient carrying the cardiac Troponin T (cTnT) mutation K273N (underwent heart transplantation) to those from “control” hearts. Preparations, mounted in a force recording apparatus (15 °C), were maximally Ca2+-activated (pCa 4.5) and fully relaxed (pCa 9) by rapid (<10 ms) solution switching. The rate constant of active tension generation following maximal Ca2+ activation (kACT) was markedly faster in the myofibrils from the K273N patient (1.7- 2 s−1) compared to controls (0.7-1 s−1). Replacement of the mutant protein by exchange with wild-type recombinant human cTn reduced kACT of HCM myofibrils close to control values (1 s−1). Force relaxation kinetics upon Ca2+ removal were also faster in K273N myofibrils than in controls, evidence that the apparent rate with which cross-bridges leave the force generating states is accelerated in the HCM preparations. The results indicate that the HCM-associated cTnT mutation K273N alters apparent cross-bridge kinetics. This can lead to increased energy cost of tension generation that may be central to the HCM disease process. Supported by the 7th Framework Program of the European Union, “BIG-HEART” grant agreement 241577, & Telethon-Italy GGP07133.I documenti in FLORE sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.