title: Changes in end-to-end interactions of tropomyosin affect mouse cardiac muscle dynamics.
abstract: The ends of striated muscle tropomyosin (TM) are integral for thin filament cooperativity, determining the cooperative unit size and regulating the affinity of TM for actin. We hypothesized that altering the alpha-TM carboxy terminal overlap end to the beta-TM counterpart would affect the amino-terminal association, which would alter the end-to-end interactions of TM molecules in the thin filament regulatory strand and affect the mechanisms of cardiac muscle contraction. To test this hypothesis, we generated transgenic (TG) mouse lines that express a mutant form of alpha-TM in which the first 275 residues are from alpha-TM and the last nine amino acids are from beta-TM (alpha-TM9aaDeltabeta). Molecular analyses show that endogenous alpha-TM mRNA and protein are nearly completely replaced with alpha-TM9aaDeltabeta. Working heart preparations data show that the rates of contraction and relaxation are reduced in alpha-TM9aaDeltabeta hearts. Left ventricular pressure and time to peak pressure are also reduced (-12% and -13%, respectively). The ratio of maximum to minimum first derivatives of change in left ventricular systolic pressure with respect to time (ratio of +dP/dt to -dP/dt, respectively) is increased, but tau is not changed significantly. Force-intracellular calcium concentration ([Ca2+]i) measurements from intact papillary fibers demonstrate that alpha-TM9aaDeltabeta TG fibers produce less force per given [Ca2+]i compared with nontransgenic fibers. Taken together, the data demonstrate that the rate of contraction is primarily affected in TM TG hearts. Protein docking studies show that in the mutant molecule, the overall carbon backbone is perturbed about 1.5 A, indicating that end-to-end interactions are altered. These results demonstrate that the localized flexibility present in the coiled-coil structures of TM isoforms is different, and that plays an important role in interacting with neighboring thin filament regulatory proteins and with differentially modulating the myofilament activation processes.
authors.affiliation: Cardiovascular Research Institute and Department of Systems Biology and Translational Medicine, College of Medicine, Texas A&M University System Health Science Center, TX 77843-1114, USA.
authors.forename: Robert D
authors.initials: RD
authors.lastname: Gaffin
authors.forename: Kuppan
authors.initials: K
authors.lastname: Gokulan
authors.forename: James C
authors.initials: JC
authors.lastname: Sacchettini
authors.forename: Timothy E
authors.initials: TE
authors.lastname: Hewett
authors.forename: Raisa
authors.initials: R
authors.lastname: Klevitsky
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Robbins
authors.forename: Vandana
authors.initials: V
authors.lastname: Sarin
authors.forename: David C
authors.initials: DC
authors.lastname: Zawieja
authors.forename: Gerald A
authors.initials: GA
authors.lastname: Meininger
authors.forename: Mariappan
authors.initials: M
authors.lastname: Muthuchamy
mesh_terms: D008841:Actin Cytoskeleton / Q000502:physiology; D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D015139:Blotting, Southern; D015153:Blotting, Western; D002118:Calcium / Q000378:metabolism / Q000502:physiology; D020013:Calcium Signaling / Q000502:physiology; D004591:Electrophoresis, Polyacrylamide Gel; D006321:Heart / Q000502:physiology*; D006339:Heart Rate / Q000502:physiology; D051379:Mice; D008822:Mice, Transgenic; D008958:Models, Molecular; D009124:Muscle Proteins / Q000378:metabolism; D009154:Mutation / Q000502:physiology; D009200:Myocardial Contraction / Q000502:physiology*; D010210:Papillary Muscles / Q000502:physiology; D011487:Protein Conformation; D015719:Single-Strand Specific DNA and RNA Endonucleases / Q000378:metabolism; D014335:Tropomyosin / Q000235:genetics / Q000502:physiology*; D016277:Ventricular Function, Left / Q000502:physiology
chemical_list: D000596:Amino Acids; D009124:Muscle Proteins; D014335:Tropomyosin; D015719:Single-Strand Specific DNA and RNA Endonucleases; D002118:Calcium