Faculty

Higher Education

M.S.in Physical Chemistry, University of Warsaw, Warsaw, Poland
Ph.D. (1989) in Natural Sciences, Nencki Institute of Experimental Biology,
Polish Academy of Sciences, Warsaw, Poland
Thesis: "The Conformational Changes of Myosin that Modulate the Interaction of the Thick and Thin Filaments of the Fast Skeletal Muscle." Supervisor - Prof. I. Kakol

Academic Experience

2006-present Associate Professor – (tenure track) in Molecular and Cellular Pharmacology Univ. of Miami, Miller School of Medicine
2006-present CV Training Grant Faculty – Mol. & Cellular Pharmacology
2003-2006 Research Associate Professor – Mol. & Cellular Pharmacology
2003-present Graduate Faculty – Mol. & Cellular Pharmacology
1997-2003 Research Assistant Professor – Mol. & Cellular Pharmacology
1995-1997 Instructor – Mol. & Cellular Pharmacology
1994-1995 Postdoctoral Associate – Mol. & Cellular Pharmacology (Dr. J. D. Potter)
1993-1994 Postdoctoral Associate – Florida State University, Institute of Molecular Biophysics, Tallahassee, FL (Dr. P. Fajer)
1990-1992 Postdoctoral Fellow – Boston Biomedical Research Institute, Boston, MA (Dr. S. Lehrer)
1986-1987 Visiting Research Fellow – Institute of Molecular Biology, Austrian Academy of Sciences, Salzburg, Austria (Dr. A. Sobieszek)
1984-1985 Visiting Research Fellow – Institute of Cytology Russian Academy of Sciences, St. Petersburg, Russia (Dr. Y. Borovikov)
1983-1990 Research Associate/Adjunct – Nencki Institute of Experimental Biology, Polish Academy of Sciences, Warsaw, Poland (Dr. I. Kakol)

Professional and Honorary Organizations

FASEB, Biophysical Society, Cardiac Muscle Society, American Heart Association.

Honors and Awards

2006-2010 NIH CCHF Study Section – standing member
2006 NSF – ad hoc reviewer
2005-2006 NIH CCHF – ad hoc reviewer
2005 Robert J. Boucek, MD Research Award – for the most meritorious research project AHA Grant-in-aid FL/Puerto Rico Affiliate
2007-2005 Grant-In-Aid 0555320B – AHA Award FL/Puerto Rico Affiliate
2004 The Alberta Heritage Foundation for Medical Research Travel Award
2004 Appreciation Award – Southern/Ohio Valley Research Consortium, Peer Review member.
2005-2004 AHA Southern/ Ohio Valley Study Section – ad hoc reviewer
2005-2003 Grant-In-Aid 0355384B - AHA Award FL/Puerto Rico Affiliate
2007-2003 NIH/NHLBI - 1R01 HL071778
2003-2001 Grant-In-Aid 0150840B AHA Award FL/ Puerto Rico Affiliate
2001-1998 Grant-In-Aid 9808237V AHA Award, FL/Puerto Rico Affiliate
1992-1991 Postdoctoral Fellowship – AHA Award Massachusetts Affiliate
1989-1987 Distinguished Scientific Achievement Award - Polish Academy of Sciences

Recent Outside Seminars

2007
Familial Hypertrophic Cardiomyopathy Mutations in Mice and Man. The Role of the Myosin Regulatory Light Chains in Cardiac Muscle Contraction- Invited external speaker for the Department of Biochemistry and Molecular Biology, Mayo Clinic, Rochester MN.

2006
Familial Hypertrophic Cardiomyopathy - The Dr. John T. MacDonald Foundation Center for Medical Genetics. Mailman Center for Child Development (MCCD) Miami

Teaching

2007 MCP 631, spring semester
2005-present Cardiovascular Journal Club for the Dept. Mol. Cellular Pharmacology Univ. of Miami School of Medicine
2001-2005 Interdisciplinary Biomedical Studies, annual lecture on Muscle Contraction, Physiology and Mechanics (Powerpoint slide presentation)
2002-present Small group sessions leader for 1 st year Medical Students in Pharmacology
2004, 2006 Training in Cardiovascular Research for Molecular and Cellular Pharmacology PhD Students
2000–1999 Biomedical Engineering Students University of Miami "Contraction of Skeletal Muscle"

Editorial responsibilities

Ad hoc reviewer for J. Biol. Chemistry, Bioph. J., Biochemistry, IUBMB Life, J. Am. Physiology, BMC Genomics, Archives of Biochemistry and Biophysics, Acta Biochimica Polonica.

Research Interest

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 the Ca²⁺ dependent 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).


Project 1

Over the past 4 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 NIH/NHLBI R01-071778


Project 2

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.

THE LABORATORY OF JAMES D. POTTER, Ph.D /
DANUTA SZCZESNA-CORDARY, Ph.D.

Pictured from left-right, front row, Yingcai Wang, Fatima deFreitas, Danuta Szczesna-Cordary (PI), James D. Potter (PI) Michelle Jones, Michelle Parvatiyar, José Renato Pinto; second row, Jiang-Sheng Liang, Zoraida Diaz-Perez, Elba Lalor, Yuhui Wen, Georgianna Guzman, Sherlley Sanon, Sanjeev Sirpal, David Dweck, Hannah Wasserman.

Personnel

Danuta Szczesna-Cordary, Ph.D. (PI)
Georgianna Guzman, Research Associate III
Michelle Jones, Research Associate III
Zoraida Diaz-Perez, Research Clinical Specialist
Yuanyuan Xu, Technical Specialist (Dr. Kerrick’s lab)
Hannah Wasserman, Intern from Columbia University, Barnard College

Postdoctoral position available

Co-Investigators at UM

Dr. James D. Potter (webpage)
Dr. W. Glenn L. Kerrick (webpage)
Dr. Yingcai Wang (webpage)

Collaborators

Dr. Jeff Moore, Boston Univ.
http://people.bu.edu/jxmoore/

Dr. Julian Borejdo, Univ. of North Texas
http://www.hsc.unt.edu/departments/molbioim/biography.cfm?id=167

Dr. Gary Lopaschuk/Cory Wagg, Univ. of Alberta
http://www.ualberta.ca/PERINATAL/Investigators/lopaschuk.htm

Dr. Ted Abraham, Johns Hopkins Univ. http://esgweb1.nts.jhu.edu/cardio/facultydtl_new.cfm?ID=126

Dr. Greg Sawicki, Univ. of Saskatchewan http://www.medicine.usask.ca/migration/pharmacology/research/grzegorz-sawicki-ph-d-1

Selected Peer-Reviewed Journal Articles

Szczesna-Cordary, D., Jones, M., Moore, J.R., Watts, J., W. Kerrick G.L., Xu, Y., Wang, Y., Wagg, C., Lopaschuk G.D. (2007) Myosin Regulatory Light Chain E22K Mutation Results in Decreased Cardiac Intracellular Calcium and Force Transients. FASEB. J. 2007 July 2; [Epub ahead of print]

Wang, Y., Szczesna-Cordary, D., Craig, R., Perez-Diaz, Z., Guzman, G., Miller, T., Potter, J.D. (2007) Fast Skeletal Muscle Regulatory Light Chain Is Required For Fast and Slow Skeletal Muscle Development. FASEB J. 21:2205-2214, 2007.

Olga M. Hernandez, Michelle Jones, Georgianna Guzman, and Danuta Szczesna-Cordary. (2007) Myosin Essential Light Chain in Health and Disease. Am J Physiol Heart Circ Physiol, 292(4):H1643-54

Wang, Y., Xu, Y., W. Kerrick, G., Wang, Y., Guzman, G., Diaz-Perez, Z., Szczesna-Cordary, D. (2006) Prolonged Ca²⁺ and Force Transients in Myosin RLC Transgenic Mouse Fibers Expressing Malignant and Benign FHC Mutations Journal of Molecular Biology, Volume 361, Issue 2, 11 August 2006, Pages 286-299.

Dumka, D. Talent, J., Akopowa, I., Guzman, G., Szczesna-Cordary, D., Borejdo, J. (2006) E22K Mutation of RLC that Causes Familial Hypertrophic Cardiomyopathy in Heterozygous Mouse Myocardium: Effect on Cross-Bridge Kinetics. Am J Physiol Heart Circ Physiol, Nov 2006; 291: H2098 - H2106. 

Hernandez, O., Szczesna-Cordary, D., Knollman, B.C., Miller, T., Bell, M., Zhao, J., Sirenko, S.G., Diaz, Z., Guzman, G., Xu. Y., Wang, Y., Kerrick, W.G., Potter, J.D. (2005) F110I and R278C Troponin T Mutations that Cause Familial Hypertrophic Cardiomyopathy Affect Muscle Contraction in Transgenic Mice and Reconstituted Human Cardiac Fibers. J. Biol. Chem. 280: 37183-37194.

Szczesna-Cordary, D., Guzman G., Zhao, J., Hernandez O., Jianqin Wei, Zoraida Diaz-Perez (2005) The E22K Mutation in Myosin RLC that Causes Familial Hypertrophic Cardiomyopathy Increases Calcium Sensitivity of Force and ATPase in Transgenic Mice. J.Cell Science 118 (16): 3675-3683.

Sawicki, G., Leon, H, Sawicka, J., Sariahmetoglu, M., Szczesna-Cordary, D., Schulz, R. (2005) Degradation of myosin light chain in isolated rat hearts subjected to ischemia-reperfusion injury: a new intracellular target for matrix metalloproteinase-2. Circulation 112: 544-552.

Szczesna-Cordary, D., Guzman G., Ng, S., Zhao, J. (2004). Familial hypertrophic cardiomyopathy-linked alterations in Ca²⁺ binding of human cardiac myosin regulatory light chain affect cardiac muscle contraction. J. Biol. Chem, 279, 3535-3542.

Szczesna-Cordary, D. (2003) Regulatory Light Chains of Striated Muscle Myosin. Structure, Function and Malfunction. Invited Review for Current Drug Targets- Cardiovascular & Hematological Disorder, 3, 1877-197.

Gomes, A.V., Potter, J.D. and Szczesna-Cordary, D. (2002) The role of Troponins in Muscle Contraction. Invited Review for IUBMB Life 54, 323-333.

Szczesna, D., Jones, M., Zhao, J., Zhu, G., Stull, J.T. and Potter, J.D. (2002) Phosphorylation of the Regulatory Light Chains of Myosin Affects Ca²⁺ Sensitivity of Skeletal Muscle Contraction. J. Applied Physiol.,92:1661-1670.

Szczesna, D. and Potter, J.D. (2002) The Role of Troponin in the Ca²⁺ Regulation of Skeletal Muscle Contraction. Results Probl Cell Differ, 36, 171-90.

Miller, T., Szczesna, D., Housmans, P.R., Zhao, J., de Freitas, F., Gomes, A.V., Culbreath, L., McCue, J., Wang, Y., Xu, Y., Kerrick, W.G. and Potter, J.D. (2001) Abnormal Contractile Function in Transgenic Mice Expressing an FHC-Linked Troponin T (I79N) Mutation. J. Biol. Chem. 276, 3743-3755.

Szczesna, D., Ghosh, D., Li, Q., Gomes, A.V., Guzman, G., Arana, C., Zhi, G., Stull, J.T., Potter, J.D.(2001) Familial hypertrophic cardiomyopathy mutations in the regulatory light chains of myosin affect their structure, Ca²⁺ binding, and phosphorylation. J. Biol. Chem. 276, 7086-7092.

Szczesna, D., Zhang, R., Zhao, J., Jones, M., Guzman, G and Potter, J.D. (2000) Altered Regulation of Cardiac Muscle Contraction by Troponin T Mutations that Cause Familial Hypertrophic Cardiomyopathy. J. Biol. Chem. 275:624-630.

Szczesna, D., Zhang, R., Zhao, J., Jones, M. and Potter, J.D. (1999) The Role of the NH2- and COOH-terminal Domains of the Inhibitory Region of TnI in the Regulation of Skeletal Muscle Contraction.  J. Biol. Chem. 274: 29536-29542.

Parsons, B., Szczesna, D., Zhao, J., Van Slooten, G., Kerrick, W.G.L., Putkey, J.A. and Potter, J.D. (1997) The Effect of pH on the Ca²⁺ Affinity of the Ca²⁺ Regulatory Sites of Skeletal and Cardiac Troponin C in Skinned Muscle Fibers. J. Muscle Res. Cell Motility. 18, 599-609.

Szczesna, D., Guzman, G., Miller, T., Zhao, J., Farokhi, K., Ellemberger, H. and Potter,J.D. (1996) The Role of the Four Ca²⁺ Binding sites of Troponin C in the Regulation of Skeletal Muscle Contraction. J. Biol. Chem. 271:8381-8386.

Szczesna, D., Zhao, J. and Potter, J.D. (1996)  Regulatory Light Chains of Myosin Modulate the Cross-Bridge Cycling in Skeletal Muscle. J. Biol. Chem., 271:5246-5250.

Szczesna, D. and Fajer, P.G. (1995) The Tropomyosin Domain is Flexible and Disordered in Reconstituted Thin Filaments. Biochemistry 34:3614-3620.

Szczesna, D., Graceffa, P., Wang, C.-L.A. and Lehrer, S.S. (1994) Myosin S1 Changes the Orientation of Caldesmon on Actin. Biochemistry 33:6716-6720.

Szczesna, D. and Lehrer, S.S. (1993) The Binding of Fluorescent Phallotoxins to Actin in Myofibrils. J. Mus. Res. Cell Mot. 14:594-597.

Szczesna, D. and Lehrer, S.S. (1992) Linear Dichroism of Acrylodan-Labeled Tropomyosin and Myosin Subfragment 1 Bound to Actin in Myofibrils. Biophys. J. 61:993-1000.

Szczesna, D., Borovikov, Y.S., Kakol I. and Sobieszek, A. (1989) Interaction of Tropomyosin with Factin-Heavy Meromyosin Complex. Biol. Chem. Hoppe-Seyler 370:399-407.

Szczesna, D., Borovikov, Y.S., Lebedeva, N.N. and Kakol, I. (1987) Effect of Phosphorylation of Myosin Light Chains on Interaction of Heavy Meromyosin with Regulated F-Actin in Ghost Fibers. Experientia 43:194-196.

Szczesna, D., Sobieszek, A. and Kakol, I. (1987) Binding of Phosphorylated and Dephosphorylated Heavy Meromyosin to F-Actin. FEBS Letters 210(2):177-180.

Kakol, I., Borovikov, Y.S., Szczesna, D., Kirillina, V.P. and Levitski, D.I. (1987) Conformational Changes of F-Actin in Myosin-Free Ghost Single Fiber Induced by either Phosphorylated or Dephosphorylated Heavy Meromyosin. Biochim. Biophys. Acta 913:1-9.

Borovikov, Y.S., Kakol, I., Szczesna, D., Kirillina, V.P. and Levitski, D.I. (1986) Effect of Phosphorylation of Rabbit Skeletal Muscle MyosinLight Chains on the Nature of Conformational Changes of F-Actin Induced by Heavy Meromyosin. Biochimia (In Russian) 51(4):691-694.

Stepkowski, D., Szczesna, D., Wrotek, M. and Kakol, I. (1985) Factors Influencing Interaction of Phosphorylated and Dephosphorylated Myosin with Actin.Biochim. Biophys. Acta 831:321-329.

Stepkowski, D., Osinska, H., Szczesna, D., Wrotek, M. and Kakol, I. (1985) Decoration of Actin Filaments with Skeletal Muscle Heavy Meromyosin Containing either Phosphorylated or Dephosphorylated Regulatory Light Chains. Biochim. Biophys. Acta 830:337-340.