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We are involved in structural studies on the assembly and function of actin-containing thin filaments in muscle and non-muscle cells. Our principal goal is to analyze and elucidate the mechanisms of thin filament-linked regulation of muscle contraction and cytoskeletal remodeling. To accomplish this goal, we use a combination of molecular biology, electron microscopy, electron tomography, image reconstruction and computational tools such as molecular dynamics protocols to better understand the interactions and dynamics of protein components of isolated and reconstituted thin filaments. Studies on mutants are carried out to elucidate abnormal filament function in disease processes.
Our laboratory was the first to directly visualize the steric-blocking mechanism of muscle regulation by identifying the positions assumed by tropomyosin on actin in the presence and the absence of Ca2+ using cryo-electron microscopy and negative staining. We also have demonstrated that during muscle activation tropomyosin moves away from myosin cross-bridge binding sites on actin in two highly cooperative steps, one induced by Ca2+ binding to troponin and a second induced by the binding of myosin to actin. Our laboratory is continuing the above-mentioned studies to obtain even greater resolution of the processes involved. At the same time, we are investigating the structural organization of troponin on thin filaments and the changes it undergoes on binding of Ca2+. We have also been engaged in studies on the structural interactions of other actin binding proteins including α-actinin, myosin binding protein-C, caldesmon, calponin, cortactin, filamin and native and mutant dystrophin, namely proteins that play important roles in the organization of the cytoskeleton in striated and smooth muscles as well as in non-muscle cells.
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Stuart Campbell,Jenette Creso, Ilayda Firlar, Saiti Halder,William Lehman,Michael Rynkiewicz,Jeffrey Moore
Journal of Cardiac Failure (2024): S3
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The Journal of general physiologyno. 5 (2024)
JOURNAL OF MOLECULAR AND CELLULAR CARDIOLOGY (2024): 30-37
The Journal of general physiologyno. 7 (2023)
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