EXCEED THE SPACE PROVIDED The thin filament regulatory proteins troponin (Tn) and tropomyosin (Tm) regulate skeletal and cardiac muscle force, the rate of shortening, and calcium sensitivity which are the ultimate expressions of the interaction between the contractile proteins myosin and actin Point, missense, and deletion mutations in troponin T and tropomyosin produce hypertrophic cardiomyopahties and alter cardiac muscle isometnc force, shortening velocity, and calcium sensitivity These changes are hypothesized to alter the cardiac function observed in patients expressing these myopathies which in turn are associated with increased incidence of heart failure and sudden cardiac death The mechanisms responsible for the changes in contractile function associated with these mutations are not understood It has been hypothesized that these proteins alter the the amount of force or displacement produced when crossbridges attach and cycle It has been hypothesized that they control the rate of cross bridge attachment, the duration of attachment, or the rate of crossbddge detachment Although recent reports have clarified the role of calcium in controlling the rate of strong cross bridge attachment, an increasing amount of data suggests that Tm and Tn also modulate maximal isometric force and unloaded shortening velocity both in normal and diseased (Hypertrophic Cardiomyopathy, HCM) muscle Further while HCM TnT and Tm mutations produce a variety of contractile defects, two common denominators in these diseases are an increased calcium sensitivity and decreased inhibition of contractility at low intracellular pH This proposars goal is to understand the mechanisms by which Tm and Tn control contractility and muscle mechanics in both health and disease The proposed experiments are designed to reveal how the three troponin subunits and tropomyosin communicate with each other and the specific aspects of contractile behavior (force, velocity of shortening, calcium sensitivity) each modifies They seek to clarify the role and mechanisms of change intracellular pH has on the mechanical behavior of the muscle and to characterize the how the activation of troponin units on the thin filament produce cooperative activation The mechanisms will be systemically examined by altering the Tn or Tm composition of regulated thin filaments and measuring contractile behavior in single molecule three bead optical trap studies, in single myofibrils, and single muscle muscle cells PERFORMANCE SITE ========================================Section End===========================================

Agency
National Institute of Health (NIH)
Institute
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Type
Research Project (R01)
Project #
5R01AR030988-23
Application #
6840823
Study Section
Special Emphasis Panel (ZRG1-SMB (01))
Program Officer
Nuckolls, Glen H
Project Start
1982-05-01
Project End
2008-01-31
Budget Start
2005-02-01
Budget End
2006-01-31
Support Year
23
Fiscal Year
2005
Total Cost
$258,869
Indirect Cost
Name
University of California Los Angeles
Department
Physiology
Type
Schools of Medicine
DUNS #
092530369
City
Los Angeles
State
CA
Country
United States
Zip Code
90095
Siththanandan, V B; Tobacman, L S; Van Gorder, N et al. (2009) Mechanical and kinetic effects of shortened tropomyosin reconstituted into myofibrils. Pflugers Arch 458:761-76
Sumandea, Marius P; Vahebi, Susan; Sumandea, C Amelia et al. (2009) Impact of cardiac troponin T N-terminal deletion and phosphorylation on myofilament function. Biochemistry 48:7722-31
Pavlov, Dmitry; Gerson, Jack H; Yu, Tianwei et al. (2003) The regulation of subtilisin-cleaved actin by tropomyosin/troponin. J Biol Chem 278:5517-22
Piroddi, N; Tesi, C; Pellegrino, M A et al. (2003) Contractile effects of the exchange of cardiac troponin for fast skeletal troponin in rabbit psoas single myofibrils. J Physiol 552:917-31
Burkart, Eileen M; Sumandea, Marius P; Kobayashi, Tomoyoshi et al. (2003) Phosphorylation or glutamic acid substitution at protein kinase C sites on cardiac troponin I differentially depress myofilament tension and shortening velocity. J Biol Chem 278:11265-72
Homsher, E; Nili, M; Chen, I Y et al. (2003) Regulatory proteins alter nucleotide binding to acto-myosin of sliding filaments in motility assays. Biophys J 85:1046-52
Tobacman, Larry S; Nihli, Mahta; Butters, Carol et al. (2002) The troponin tail domain promotes a conformational state of the thin filament that suppresses myosin activity. J Biol Chem 277:27636-42
Karibe, A; Tobacman, L S; Strand, J et al. (2001) Hypertrophic cardiomyopathy caused by a novel alpha-tropomyosin mutation (V95A) is associated with mild cardiac phenotype, abnormal calcium binding to troponin, abnormal myosin cycling, and poor prognosis. Circulation 103:65-71
Strand, J; Nili, M; Homsher, E et al. (2001) Modulation of myosin function by isoform-specific properties of Saccharomyces cerevisiae and muscle tropomyosins. J Biol Chem 276:34832-9
Gordon, A M; Regnier, M; Homsher, E (2001) Skeletal and cardiac muscle contractile activation: tropomyosin ""rocks and rolls"". News Physiol Sci 16:49-55

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