Boston, MA, **/**/****

Cover letter

Pawel T. Szymanski, Ph.D.

To whom it may concern,

Currently, I am seeking Associate Scientist I Position in Technical Development (job ID

#**-*****).

In 2010, I accepted Research Associate Position in the Department of

Pharmaceutical Sciences at South Dakota State University in Brookings. When I made

the transition to SDSU, my aim was to gain more knowledge about transitional research

in regards to searching for an anti-cancer drug(s). Prior to this career switch, I was a

traditional protein biochemist dealing with both contractile and cytoskeleton sub-domains

in smooth muscle.

Another reason I moved to a more applied science and started dealing with

tumor cells was because of the far reaching similarities between smooth muscle and the

non-muscle contractile systems.

As you can see in my CV, I am a versatile biochemist possessing a solid

publication record and good oral and written communication skills. I have extensive

laboratory experience within academic settings, and a proven track record of successfully

managing multiple research projects in the interdisciplinary settings.

I would certainly welcome the opportunity to talk to you about the position,

should you find my background suitable for your program. I appreciate your time, and I

look forward to hearing from you. Enclosed you will find my CV for your review, which

includes a bibliography of my publications and a list of my personal references.

Additional information is available upon request.

Sincerely yours,

Dr. Pawel T. Szymanski

SUMMARY

So far, my research has been focus on regulation of the actin-myosin interaction

and cytoskeleton remodeling in smooth muscle, tissue that shows a high degree similarity

with the non-muscle cells, including tumor cells. Until now, I specialized in signaling

pathways involving Ca2+, Ca2+ /calmodulin (CaM), myosin light chain kinase (MLCK),

protein kinase C (PKC), c-AMP protein kinase, tyrosine kinases and phosphatases type 1

and type 2A.

While performing in the in vitro studies, I purified and characterized many

proteins involved in contractility, including actin, CaM, calponin (CaP), troponin-I (Tn-

I), myosin and tropomyosin, MLCK and protein phosphatases (type 1 and 2A), filamin,

desmin and the Arp2/3 complex. I also extracted and then reconstituted CaP, caldesmon,

myosin regulatory light chain and SM22 from/into chemically skinned smooth muscle

fibers. I have hands-on experience in both traditional ion-exchange chromatography and

gel filtration. I also applied gel filtration on Superose-12 using an Akta program to run

HPLC. I also used an affinity chromatography to purify native and tagged proteins. I am

knowledgeable in all kinds of electrophoresis, including 2D electrophoresis, native

electrophoresis, IEF-urea gel separation, Western blot and blot overlay assay with the use

of the hemagglution-tagged CaM, zymography and immunoprecipitation. I also have

experience in co-sedimentation assays and labeling proteins with fluorescently sensitive

dyes (IAE-DANS-CaP to mimic phosphorylation and pyrene-actin) to determine binding

parameters of interaction, viz., affinity and stochiometry in solution using fluorescence

intensity measurements. I am also versed in falling-ball assay, light-scattering and

tyrosine and tryptophan intrinsic fluorescence to determine conformational changes

within proteins. I utilized various proteases to localize protein regions involved in

protein-protein interaction and viscosity measurement to determine the state of assembly

of both actin and myosin filaments.

I have experience in many analytical methods, such as cell fractionation, thyroid

hormones RIA, radioiodination of the low molecular mass molecules, Scatchard analysis

and determination of activities of phospholipid-synthesizing and phospholipid-

degradation enzymes.

I also gained some experience in gene cloning, plasmid transformation and

bacteria expression of αCaP and yeast tropomyosin isoform-2 (TPM-2).

Quite recently, I gained substantial knowledge in cell culture, the Elisa-type assay

to characterize and better understanding the anti-cancer activity of sarcophine-diol, a

semi-synthetic derivative of sarcophine. These assays include application of Trypan-blue

staining, MTT, BrdU, TACS Apoptotic DNA Laddering, Poly (ADP-ribose) polymerase,

and phospholipases A2 and C enzymatic kits. I also determine the expression levels of

several pro-cancer and anti-apoptotic genes in melanoma cells treated with many

different potential anti-cancer natural products from the Red Sea soft coral that I used as

a model for tumor cells, and compared to those in normal healthy monkey kidney cells.

I am enclosing a list of my publications to prove my biochemical skills.

CURRICULUM VITAE

Pawel T. Szymanski, Ph.D.

30 Taft St., Apt. # 3

Boston, MA 02125

Cell phone # 617-***-****

Personal e-mail: acpykl@r.postjobfree.com

OBJECTIVE

Currently, I am seeking Associate Scientist I Position in Technical Development (job ID

#15-01021).

I am a versatile protein biochemist with a solid bench-working experience in the

fields of protein-protein interaction and signaling pathways as it relates to structure-

function relationships and protein folding/unfolding within cytoskeleton domain in

smooth muscle, a tissue that shows a high degree similarity with the non-muscle cells,

including tumor cells. These similarities pertain to the protein contents and isoform

composition of cytoskeleton and signaling transduction pathways involved in

contractility in smooth muscle and rapid, uncontrolled division in cancer cells.

FORMAL EDUCATION

1979-87 Ph.D., Biochemistry, Medical Research Center, Warsaw, Polish Academy of

Science (“The effects of thyroid hormones on 1-adrenergic receptors in the

rat heart muscle”) experiments were performed in Medical Center for

Postgraduate Education, Warsaw, Poland, promotor: Dr. Andrzej Gardas.

M.Sc., Biochemistry, University of Warsaw, Poland (“Characterization of

1974-79

polyprenols in Calendula officinalis L promotor: Dr. Zofia Kasprzyk.

1972-77 M.Sc., Animal Engineering, Agriculture University, Warsaw, Poland

(“Fermentation of pectin by pure strains and combined cultures of rumen

bacteria”), experiments were performed in the Institute of Animal Nutrition,

Polish Academy of Science, Jablonna/near Warsaw, Poland, supervisor: Dr.

Alexander Ziolecki, promotor: Dr. Jolanta Zawitkowska.

ADDITIONAL TRAINING

1998 Emerging Strategies for Management of Upper Gastrointestinal Motility

Disorders (Rush-Presbyterian-St.Luke's Medical Center, New Orleans, LA).

1985 Advanced Course on Thyroid Hormones Radioimmunoassay (Medical Center

for Postgraduate Education, Warsaw, Poland).

1984 International School on Biomembranes and Receptors Mechanisms (Katania,

Sicily, Italy).

1983 FEBS/SKMB/SNG Advanced Course on Enzymes, Receptors and Carrier in

Biological Membranes (Gwatt and University of Bern, Switzerland).

POSITIONS HELD

2010-2012 Research Associate, Department of Pharmaceutical Sciences, South

Dakota State University, Brookings, SD.

Dec 2009 - May 2010: Research Associate, Department of Physiology, Wayne State

University, Detroit, MI.

2005 Post-doctoral Fellow, Department of Cell Biology, Harvard Medical

School, and Research Specialist at Stowers Institute for Medical Research,

Kansas City, MO.

Research Fellow, Division of Hematology, Brigham and Women’s

2004

Hospital, Harvard Medical School, Boston, MA.

2000-03 Research Associate, Physiology Program, Department of Environmental

Health, Harvard School of Public Health, Boston, MA.

1996-2000 Research Associate, Center for Swallowing and Motility Disorders,

Veteran’s Administration Medical Center, Harvard Medical School, West

Roxbury, MA.

1992-96 Research Associate, Muscle Research Group, Boston Biomedical

Research Institute, Boston, MA.

1988-92 Post-doctoral Fellow, Department of Physiology and Biophysics,

University of Cincinnati, College of Medicine, Cincinnati, OH.

BIBLIOGRAPHY

Original Publications:

1. Szymanski PT. A note on the fermentation of pectin by pure strains and

combined cultures of rumen bacteria. Acta Microbiol Polon 1981;30:159-63.

Szymanski PT, Nauman J. -adrenergic receptors in the heart muscle of the rat.

2.

Optimal conditions for interaction of 125IHYP with -adrenergic receptors.

Endokrynol Pol 1983;34:253-66.

Szymanski PT. -adrenergic receptors in the heart muscle of the rat. Kinetic

3.

parameters of interaction of 125IHYP with -adrenergic receptors. Endokrynol Pol

1983;34:305-14.

4. Szymanski PT, Nauman J. Effects of thyroid hormones and reverse

triiodothyronine pretreatment on -adrenergic receptors in the rat heart. Acta

Physiol Polon 1986;37:131-8.

Szymanski PT. Effect of glycoprotein modification on -adrenoreceptors binding

5.

ability in the rat cardiac membranes. Acta Physiol Polon 1987;38:483-92.

6. Famulski KS, Szymanska G, Szymanski P, Sarzala MG. Hyperthyroidism affects

the activity of cAMP-dependent protein kinase and protein phosphorylation in the

heart sarcolemma. Biomed Biochem Acta 1987;46:448-51.

7. Szymanski PT, Szymanska G, Pikula S, Sarzala MG. The effects of thyroid

hormones on lipid composition and activities of lipid synthesizing enzymes in the

rat heart sarcolemma. Endokrynol Pol 1989;40:43-9.

8. Strauss JD, Szymanski PT, DiSalvo J, Paul RJ. Effects of modulators of myosin

phosphorylation on isometric force and shortening velocity in skinned smooth

muscle. In: Sperlakis N, Wood JD, eds., Frontiers in Smooth Muscle Research.

New York: Alan R. Liss, Inc., 1990;617-22.

9. Rymaszewski Z, Szymanski PT, Abplanalp WA, Myatt L, DiSalvo J, Cohen RA.

Human retinal vascular cells differ from umbilical cells in synthetic functions and

their response to glucose. Proc Soc Exp Biol Med 1991;199:183-91.

10. Szymanski PT, Paul RJ. Polylysine: an activator of smooth muscle contractility.

Adv Exp Med Biol 1991;304:363-8.

11. Szymanski PT, Strauss JD, Doerman G, DiSalvo J, Paul RJ. Polylysine activates

smooth muscle actin-myosin interaction without LC20 phosphorylation. Am J

Physiol 1992;262 (Cell Physiol. 31):1446-55.

12. Szymanski PT, Ferguson DG, Paul RJ. Polylysine activates smooth muscle

myosin ATPase activity via induction of a 10S-6S transition. Am J Physiol

1993;265 (Cell Physiol. 34):379-86.

13. Szymanski PT, Tao T. Interaction between calponin and smooth muscle myosin.

FEBS Lett 1993;331:256-9.

14. Obara K, Szymanski PT, Tao, Paul RJ. The effects of calponin on isometric force

and shortening velocity in permeabilized taenia coli smooth muscle. Am J Physiol

1996;270 (Cell Physiol. 39):481-7.

15. Szymanski PT, Grabarek Z, Tao T. Correlation between calponin and myosin

subfragment 1 binding to F-actin and ATPase inhibition. Biochem J

1997;321:519-23.

16. Szymanski PT, Tao T. Localization of protein regions involved in the interaction

between calponin and myosin. J Biol Chem 1997;272:11142-6.

17. Hirano I, Kakkar R, Saha JK, Szymanski PT, Goyal RK. Tyrosine

phosphorylation in contraction of opossum esophageal longitudinal muscle in

response to SNP. Am J Physiol 1997;273 (Cell Physiol. 36):247-52.

18. Tang JX, Szymanski PT, Janmey PA, Tao T. Electrostatic effects of smooth

muscle calponin on actin assembly. Eur J Biochem 1997;247:432-40.

19. Szymanski PT, Chacko TK, Rovner AS, Goyal RK. Differences in contractile

protein contents and isoforms in phasic and tonic smooth muscles. Am J Physiol

1998;275 (Cell Physiol. 44) 684-92.

20. Szymanski PT, Goyal RK. Calponin binds to the 20-kilodalton regulatory light

chain of myosin. Biochemistry 1999;38:37778-84.

21. Szymanski PT, Szymanska G, Goyal RK. Differences in calmodulin and

calmodulin-binding proteins in phasic and tonic smooth muscles. Am J Physiol

2002;282; (Cell Physiol. 48) 94-104.

22. Szymanski PT, Ferguson DG, Paul RJ. Polylysine binding to unphosphorylated

smooth muscle myosin enhances formation and stabilizes myosin filaments in

vitro. Acta Physiol Scand 2002;174:337-46.

23. Szymanski PT, Dickie R, Rogers R, Fredberg JJ. Extraction and reconstitution of

calponin and consequent contractile ability in permeabilized smooth muscle

fibers. Anal Biochem 2003;321;8-21.

Nakamura F, Stossel TP, Hartwig JH, Szymanski PT. Ca2+ and calmodulin

24.

regulate the binding of filamin A to actin filaments. J Biol Chem 2005;280:32426-

33.

25. Szymanski PT, Kuppast B, Ahmed SA, Khalifa S, Fahmy H. Sarcophine-diol, a

skin cancer chemopreventive agent, inhibits proliferation and stimulates apoptosis

in mouse melanoma B16F10 cell line. Mar Drugs (2012) 10:1-19.

26. Szymanski PT, Muley P, Ahmed SA, Khalifa S, Fahmy H. Sarcophine-diol

inhibits expression of Cox-2, inhibits activity of cPLA2, enhances degradation of

PLA2 and PLC 1 and inhibits cell membrane permeability in mouse melanoma

B16F10 cells. Mar Drugs (2012) 10:2166-2180.

27. Abou El-Ezz RF, Ahmed SA, Radwan MM, Ayoub NA, Afifi MS, Ross, SA,

Szymanski PT, Fahmy H, Khalifa SI. Biactive cembranoids from the Red Sea

coral Sarcophyton glaucum.Tetrahedron Lett (2013) 54:989-992.

28. Szymanski PT, Ahmed SA, Khalifa S, Tokuda H, Ichiishi E, Iida A, Suzuki N,

Fahmy H. Chemopreventive effect of sarcophine-diol on NOR-1 induced TPA

promoted skin cancer carcinogenesis in female HOS:HR-1 mice. Nat Prod

Commun (2013) 8:153-154.

29. Szymanski PT, Ahmed SA, Radwan MM, Khalifa SI, Fahmy H. Evaluation of

the anti-melanoma activities of sarcophine, -7 ,8 -

dihydroxydeepoxysarcophine and sarcophytolide from the Red Sea soft coral

Sarcophyton glaucum. Nat Prod Commun (2014) 9:151-154.

30. Szymanski PT, Ahmed SA, Radwan MM, Khalifa SI, Fahm y H. Evaluation of

the anti-melonoma activities of (1S,2E,4R,6E,8R,11S,12R)-8,12 epoxy-2,6

cembradiene-4,11-diol, (1S,2E,4R,6E,8S,11R,12S)-8,11 epoxy 4,12 epoxy-2,6-

cembradiene and (1S,4R,13S)-cembra-2E,7E,11E-trien-4,13-diol from the Red

Sea soft coral Sarcophyton glaucum. Nat Prod Commun (2014) 9:1143-1146.

31. Szymanski PT, Philippe J, Kuppast B, Ahmed SA, Khalifa S, Fahmy H.

Sarcophine-diol increases degradation of nucleic acids and modulates expression

of genes involved in the anti-proliferative and pro-apoptotic cascades in mouse

melanoma B16F10 cell line. (Manuscript in preparation for submission).

Review Articles:

1. Szymanski PT. Adrenergic receptors. Post Biochem 1983:67:91-115.

2. Szymanski PT. Electrophoresis. In: Gardas A, ed. New methods in protein

purification. Warsaw: Medical Center of Postgraduate Education, Handbook for

Students 1985:75-91.

Szymanski PT. Thyroid hormones and -adrenergic receptors. Kardiol Pol

3.

1988;31:662-84.

Szymanski PT. Calponin (CaP), as a latch-bridge protein – a new concept in

4.

regulation of contractility in smooth muscles. J Muscle Res Cell Motil 2004;25:7-

19.

5. Szymanski PT. Protein assembly and regulation of contractility in striated

muscles (fast and slow twitched and cardiac muscles), smooth muscle and non-

muscle tissues – similarities and differences. (Manuscript in preparation for

submission).

REFERENCES:

Dr. Hesham Fahmy

Associate Professor

Department of Pharmaceutical Sciences

College of Pharmacy

South Dakota State University

One Administration Lane

Brookings, SD, 57007

phone 605-***-****

e-mail: acpykl@r.postjobfree.com

Dr. Jeffrey J. Fredberg

Professor

Physiology Program

Department of Environmental Health

Harvard School of Public Health

665 Huntington Avenue

Boston, MA 02115

phone 617-***-****

e-mail: acpykl@r.postjobfree.com

Dr. Raj K. Goyal

Emeritus Professor

Veteran’s Affair Medical Center

Harvard Medical School

1400 VFW Parkway

West Roxbury, MA 02132

e-mail: acpykl@r.postjobfree.com

Dr. Richard J. Paul

Emeritus Professor

Department of Molecular and Cellular Physiology

University of Cincinnati, College of Medicine

231 Albert Sabin Way

Cincinnati, OH 45267-0576

e-mail: acpykl@r.postjobfree.com

Dr. Thomas P. Stossel

Professor

Translational Medicine Division

Hematology Division

Brigham and Women’s Hospital

Harvard Medical School

One Blackfan Circle

Boston, MA 02115

phone 617-***-****

e-mail: acpykl@r.postjobfree.com

Dr. Terence Tao

Retired Senior Scientist

Muscle Research Group

Boston Biomedical Research Institute

64 Grove Street

Watertown, MA 02472

e-mail: acpykl@r.postjobfree.com

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