Danuta Szczesna-Cordary, Ph.D.
Professor of Molecular & Cellular Pharmacology
Rosenstiel Medical Sciences Building 6113
2012 pres Professor of Molecular and Cellular Pharmacology (with tenure) University of Miami Miller School of Medicine
2011 pres Director, Training Program in Cardiovascular Signaling
2006 2012 Associate Professor
2003 2006 Research Associate Professor
1997 2003 Research Assistant Professor
Honors and Awards
2010 Award of Tenure
2010 Appreciation of Services Award, Center for Scientific Review National Institutes of Health
2005 Robert J. Boucek, MD Research Award FL/Puerto Rico Affiliate
2004 Certificate of Appreciation, American Heart Association
2004 The Alberta Heritage Foundation for Medical Research Travel Award
1989 Distinguished Scientific Achievement Award Polish Academy of Sciences
2012 pres NIH/NIAMS Skeletal Muscle and Exercise Physiology Study Section (SMEP), member
2005 2010 NIH/NHLBI Cardiac Contractility Hypertrophy and Failure (CCHF), member
2006 NSF, ad hoc reviewer
2004 2005 AHA Southern/Ohio Valley, panel reviewer
Univ. of Miami Miller School of Medicine
1600 NW 10th Ave., R-189, 6113 RMSB
Miami, FL 33136
THE SZCZESNA-CORDARY LAB
Pictured from left-to-right: Stephen Ryan - UM Intern, Wenrui Huang - 6th year PhD Student (Graduated in January 2015), Chen-Ching (Vicky) Yuan 3rd Year PhD Student, Hanyao (Brandon) Foong 2nd Year MD Student, Danuta Szczesna-Cordary, PhD - PI, Ana I. Rojas, BS - Research Associate II, Zhiqun (Cindy) Zhou, PhD - Postdoctoral Associate, Katarzyna Kazmierczak, PhD - Assistant Scientist, and Jingsheng Liang, MD - Associate Scientist
The major area of research in my laboratory is related to the Ca2+ regulation of striated (skeletal and cardiac) muscle contraction. In particular, we focus on the role of the myosin regulatory (RLC) and essential (ELC) light chains in force generation and the kinetics of myosin cross-bridges.
Figure 1. The myosin head (S1) derived from the atomic structure by Rayment et al., 1993 . The heavy chain is shown in green, red, and blue to highlight functional domains. The essential light chain and regulatory light chain function to support the myosin neck region and are colored in yellow ( ELC) and magenta ( RLC).
There are three major and functionally different domains in the myosin molecule: a motor domain, a lever arm domain - both located in the myosin head (S1) (Fig. 1), and a tail region (Rayment et al., 1993; Saraswat and Lowey, 1998; Waller et al., 1995; Xie et al., 1994) . The myosin motor domain contains a catalytic site, also called an ATP binding pocket, and an actin binding domain. A small converter domain links the myosin motor domain to the lever arm region (Fig. 1). The lever arm domain of muscle myosin is composed of a long helix containing two IQ motifs (IQxxxRGxxxR) that form the attachment sites for the ELC and the RLC (Dominguez et al., 1998; Houdusse et al., 1999; Rayment et al., 1993; Saraswat and Lowey, 1998; Xie et al., 1994) (Fig. 1). The physiological importance of these two myosin light chains has been recently highlighted by the discovery of genetic mutations shown to cause Familial Hypertrophic Cardiomyopathy (FHC) (Fig. 2).
Figure 2. The regulatory domain of scallop myosin
(1WDC) by Houdusse and Cohen, 1996 .
The heavy chain (MHC) is shown in blue, the essential
light chain (ELC) in yellow and the regulatory light chain
(RLC) in red. The FHC mutations in ELC and RLC are
indicated with arrows.
Cardiovascular diseases are the number one cause of mortality worldwide with heart failure being highly prevalent in most affluent parts of the world. There is an urgent need for a better understanding of the mechanisms underlying FHC that often leads to premature sudden cardiac death (SCD) . Our research addresses the mechanisms by which mutations in myosin regulatory light chain and essential light chain cause FHC and lead to SCD (Fig. 2).
Over the past 8 years our laboratory has been studying the functional consequences of several FHC RLC mutations (E22K, N47K and R58Q) expressed in transgenic mice (Fig. 3). Our recent results revealed new possible mechanisms by which the FHC-mutated RLC proteins may affect the contractile function of the mutated myocardium. One of the mechanisms is based on the hypothesis that in healthy muscle, the RLC functions as a temporary intracellular Ca2+-buffer working in parallel with the sarcoplasmic reticulum (SR) Ca2+ pump in sequestering Ca2+, thereby promoting muscle relaxation. We hypothesize that by changing the properties of the RLC Ca2+-Mg2+ binding site (Fig. 3), the FHC mutations can facilitate or inhibit this intracellular RLC function and result in increased or decreased kinetics of muscle relaxation. Another hypothesis pertains to the mutation controlled metal occupancy of the Ca2+-Mg2+ binding site of RLC and the mechanism by which Ca2+ or Mg2+ binding to RLC may influence the interaction of myosin with actin and tension generation.
During muscle contraction, the increase in Ca2+ concentration activates the Ca2+-calmodulin dependent myosin light chain kinase (MLCK) and leads to phosphorylation of the RLC (at serine 15). Our solution studies showed a link between the effect of the specific FHC mutation and RLC phosphorylation and implicated both events, the Ca2+ binding to the RLC and its MLCK-phosphorylation play a key role in the regulation of cardiac muscle contraction. Both of these processes, most likely, operate as adaptive and/or protective mechanisms to either attenuate the effect of the FHC mutations and/or improve performance of the working muscle. Based on our findings and those of others, we further hypothesize that an FHC induced pathological cardiac phenotype can be rescued by Ca2+-calmodulin activated MLCK phosphorylation of the RLC-mutated myocardium. The specific questions that we ask are:
Figure 3. The regulatory light chain of myosin derived from the atomic
structure of S1 (2MYS) by Rayment et al., 1993 . The Ca2+-binding loop is
shown in green. The sites of FHC mutations are pointed with arrows.
- Do FHC induced changes in the properties of the RLC Ca2+-Mg2+ binding site inhibit or facilitate the function of RLC as a temporary intracellular calcium buffer? Do FHC mutations shift the metal occupancy of the RLC Ca2+-Mg2+ binding site during muscle contraction?
- Is RLC phosphorylation by Ca2+-calmodulin ( CaM) activated myosin light chain kinase (MLCK) affected by FHC-linked RLC mutations? Can MLCK phosphorylation rescue a mutation induced pathological cardiac phenotype?
- Do FHC-associated mutations in RLC alter intermolecular interactions between RLC and myosin heavy chain (HC) and ultimately myosin and actin? Do these changes lead to myofilament disarray, cardiac hypertrophy and dysfunction of the mutated myocardium?
This area of our research is funded by 5R01HL071778-07 and 5R01HL090786-02.
This project focuses on two areas of research:
- The importance of the direct N-terminal interaction of the ventricular myosin ELC with actin during cardiac muscle contraction.
- The mechanisms by which FHC mutations in ELC alter the physiological properties of cardiac muscle.
Transgenic mice have been generated expressing various levels of a truncated ventricular ELC, lacking 43 amino acids from its N-terminus (ELC-D 43), to mimic the distribution of the ELC observed in fast skeletal muscle. As a result, the mouse cardiac muscle contains varying ratios of the long (endogenous) to short (transgenic) ELC forms. Our hypothesis is that increasing ratios of short transgenic ELC- D 43 to long endogenous ELC will result in a progressively weakened binding of myosin to actin, decreased force development and ultimately in compromised cardiac muscle performance. We are testing this hypothesis by measuring the actin-myosin interaction and force development at the single molecule level and in skinned and intact muscle fibers from transgenic ELC- D 43 mice.
In addition, we are studying the mechanisms by which FHC mutations in ELC alter the physiological properties of cardiac muscle. To date five mutations in the MYL3 gene that encodes the human ventricular ELC have been associated with FHC (E56G, A57G, E143K, M149V, R154H). Various mutation-specific phenotypes in humans including multiple cases of SCD at a young age have been observed. Transgenic mice expressing these FHC ELC mutations have been generated and the physiological consequences studied in skinned and intact muscle fibers in vitro and in vivo by echocardiography, MRI and hemodynamic methods. Our hypothesis is that FHC ELC mutations will affect the mechanical and enzymatic properties of myosin by altering the interaction of the ELC with the heavy chain of myosin and/or the interaction of the N-terminus of ELC with actin and consequently lead to a compromised interaction of myosin with actin in the mutated myocardium. The most profound changes are expected with those FHC mutations that result in poor prognosis and SCD in humans.
Co-Investigators at UM
Dr. Julian Borejdo, Univ. of North Texas
Our collaborative research is funded by NIH/NHLBI R01-090786
Dr. Greg Sawicki, Univ. of Saskatchewan
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Phosphorylation of Cardiac Myosin Regulatory Light Chain Prevents Development
of Hypertrophic Cardiomyopathy in Mice. Yuan CC, Muthu P, Kazmierczak K, Liang
J, Huang W, Irving TC; Kanashiro-Takeuchi RM, Hare JM, and Szczesna-Cordary
D. Proc Natl Acad Sci U S A. 2015 Jun 29. pii:201505819. [Epub ahead of print] PMID: 26124132
Karabina A, Kazmierczak K, Szczesna-Cordary D, Moore JR. Arch Biochem Biophys. 2015 Jun 25. pii: S0003-9861(15)00278-7. doi: 10.1016/j.abb.2015.06.014. [Epub ahead of print] PMID: 26116789
Huang. W., Liang, J., Kazmierczak, K., Muthu, P., Duggal, D., Farman, G.P., Sorensen, L., Pozios, I., Abraham, T., Moore, J.R., Borejdo, J., and Szczesna-Cordary, D. (2014) Hypertrophic Cardiomyopathy Associated Lys104Glu Mutation in the Myosin Regulatory Light Chain Causes Diastolic Disturbance in Mice
. J Mol Cell Cardiol. 74:318-329. PMID: 24992035
Kazmierczak, K., Yuan, C-C., Liang, J., Huang, W., Rojas, A.I. and Szczesna-Cordary, D. (2014) Remodeling of the heart in hypertrophy in animal models with myosin essential light chain mutations
. Front. Physiol. 22 September 2014 doi: 10.3389/fphys.2014.00353
Muthu P, Liang J, Schmidt W, Moore JR, Szczesna-Cordary D. (2013) In Vitro Rescue Study of a Malignant Familial Hypertrophic Cardiomyopathy Phenotype by Pseudo-Phosphorylation of Myosin Regulatory Light Chain. Arch Biochem Biophys. 2013 Dec 26. pii: S0003-9861(13)00385-8. doi:
10.1016/j.abb.2013.12.011. [Epub ahead of print] PMID: 24374283 [PubMed - as supplied by publisher] Related citations
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