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Custom Vector Construction> | Purification of recombinant rat cardiac Tn subunits Recombinant c-myc tagged TnT variants (WT, K211D, K211D ? T204E, R206W, R206W ? T204E), rat cardiac troponin I (TnI), and rat cardiac troponin C (TnC) were cloned into pSBETa vector (GenScript Corporation, Piscataway, NJ) and expressed in BL21*DE3 cells (Novagen, Madison, WI), as described previously (Chandra et al. | Get A Quote |
Failing hearts of dilated cardiomyopathy (DCM)-patients reveal systolic dysfunction and upregulation of several Protein Kinase C (PKC) isoforms. Recently, we demonstrated that the functional effects of T204E, a PKC phosphomimic of cardiac troponin T (TnT), were differently modulated by α- and β-myosin heavy chain (MHC) isoforms. Therefore, we hypothesized that the interplay between the effects of T204E and a DCM-linked mutation (K211Δ or R206W) in TnT would modulate contractile parameters linked-to systolic function in an MHC-dependent manner. To test our hypothesis, five TnT variants (wildtype, K211Δ, K211Δ + T204E, R206W, and R206W + T204E) were generated and individually reconstituted into demembra... More