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A. Statement B. Positions and Honors BIOGRAPHICAL SKETCH NAME POSITION TITLE Heuck, Alejandro Pablo Associate Professor of Biochemistry and Molecular Biology eRA COMMONS USER NAME AHEUCK EDUCATION/TRAINING DEGREE INSTITUTION AND LOCATION YEAR(s) FIELD OF STUDY (if applicable) University of Buenos Aires, Argentina B.S. 1983 – 1991 Chemistry University of Buenos Aires, Argentina Ph.D. 1992 - 1997 Biochemistry Texas A&M University, College Station, TX Post-doc 1998-2001 Biochemistry A. Statement I have a broad background in biophysics, with specific training and expertise in protein-membrane interactions, pore-forming toxins, and fluorescence spectroscopy required for this application. As a postdoctoral fellow at Texas A&M University, I carried out structural research and functional analysis on cholesterol-dependent pore forming proteins. As an Assistant Professor of Biochemistry and Molecular Biology I have initiated two independent projects in the area of protein-membrane interactions which have been funded mainly by the American Heart Association, the American Lung Association, and NIH. The first involves the development of molecular probes to detect cholesterol levels in cell membranes. The second project is focused in the functional and structural analysis of the Pseudomonas aeruginosa type three secretion translocon complex, by which toxins are directly injected into target cells. B. Positions and Honors. Positions 1992-1997 Doctoral Student, (with Ricardo Wolosiuk) Instituto Leloir, University of Buenos Aires, Argentina. 1997-2001 Post Doctoral Research Associate, (with Arthur E. Johnson) Texas A&M University System Health Science Center. 2001-2005 Senior Research Associate, (with Arthur E. Johnson) Texas A&M University System Health Science Center. 2005-2013 Assistant Professor, University of Massachusetts, Amherst. Dept. Biochemistry and Molecular Biology 2013-present Associate Professor, University of Massachusetts, Amherst. Dept. Biochemistry and Molecular Biology Honors 1992-1993 Junior Fellowship Award for Graduate Research, CONICET, Argentina. 1994-1995 Senior Fellowship Award for Graduate Research, CONICET, Argentina. 1996 Fellowship for Graduate Research, Instituto Leloir Foundation, Argentina. 2002 Distinguished Poster Presentation Award FASEB Summer Conferences, Molecular Biophysics of Cellular Membranes, (Vermont, USA). 2009-2010 American Lung Association Scholar: Diseases of Infants and Children 2011 John Adams Investigator, named by the Massachusetts Technology Collaborative’s John Adams Innovation institute. Other Experience and Professional Service Journal and Grant Review Service Nature Chemical Biology Current Protein & Peptide Science PLoS Pathogens Journal of Experimental Medicine The Journal of Biological Chemistry Protein Science Biochemistry(ACS) FEBS Letters Biophysical Journal Journal of Medicinal Chemistry BMC Microbiology International Journal of Molecular Sciences Protein Science Toxins FEMS Microbiology Letters Open Biology BBA Biomembranes The Wellcome Trust, England (Molecular and Biophysical Chemistry Cellular Biology) PLoS One Faculty Research/Healey Endowment Grant Journal of Molecular Biology University of Massachusetts Amherst. Professional Associations 2005-present Chemistry & Biology Interface Training Program, UMass Training Faculty. 2005-present Molecular and Cellular Biology Program, UMass Faculty Member. 2009-present Institute for Cellular Engineering, UMass Training Faculty. 2009-present Biophysical Society 2009-present Institute for Drug Resistance. Member C. Selected peer-reviewed publications (in chronological order). Heuck A.P., Wolosiuk R.A. (1997) Di-fluoresceintiocarbamyl-insulin: A Fluorescent Substrate for the Assay of Protein Disulfide Oxidoreductase Activity. Analytical Biochemistry 248, 94-101. PMID 9177728 Shatursky O.*, Heuck A.P.*, Shepard L.A., Rossjohn J., Parker M.W., Johnson A.E. and Tweten R.K. (1999) The Mechanism of Membrane Insertion for a Cholesterol-Dependent Cytolysin: A Novel Paradigm for Pore-Forming Toxins. Cell, 99, 293-299. *These authors contributed equally to this work. PMID 10555145 Heuck A.P., Hotze E.M., Tweten R.K., and Johnson A.E. (2000) Mechanism of Membrane Insertion of Multimeric - Barrel Protein: Perfringolysin O Creates a Pore Using Ordered and Coupled Conformational Changes. Molecular Cell, 6, 1233-1242. PMID 11106760 Heuck A.P., Tweten R.K., and Johnson A.E. (2001) -Barrel Pore-Forming Toxins: Intriguing Dimorphic Proteins Biochemistry 40, 9065-9073. PMID 11478872 Ramachandran R., Heuck A.P., Tweten R.K., and Johnson A.E. (2002) Structural Insights into the mechanism of a cholesterol-dependent cytolysin Nature Structural Biology 9, 823-7. PMID 12368903 Heuck A.P., Tweten R.K., Johnson A.E. (2003) Assembly and Topography of the Prepore Complex in Cholesterol- Dependent Cytolysins. Journal of Biological Chemistry 278, 31218-31225. PMID 12777381 Heuck A.P.*, Savva C.G., Holzenburg A., Johnson A.E. (2007) Conformational changes that effect oligomerization and initiate pore formation are triggered throughout perfringolysin O upon binding to cholesterol. Journal of Biological Chemistry 282, 22629-22637. *Corresponding author. PMID 17553799 Flanagan J.J., Tweten R.K., Johnson A.E., Heuck A.P. (2009) Cholesterol Exposure at the Membrane Surface is Necessary and Sufficient to Trigger Perfringolysin O Binding. Biochemistry, 48, 3977-3987. PMID 19292457 Moe P.C., Heuck A.P. (2010) Phospholipid Hydrolysis Caused by Clostridium perfringens -toxin Facilitates the Targeting of Perfringolysin O to Membrane Bilayers. Biochemistry, 49, 9498-9507. PMID 20886855 Raghava S., Giorda K. M., Romano F. B., Heuck A. P, Hebert D. (2011). The SV40 late protein VP4 is a viroporin that forms pores to disrupt membranes for viral release. PLoS Pathogens, 7, e1002116. PMID: 21738474 Romano FB, Rossi KC, Sava CG, Holzenburg A, Clerico EM, Heuck AP. (2011). Efficient isolation of Pseudomonas aeruginosa type III secretion translocators and assembly of heteromeric transmembrane pores in model membranes. Biochemistry, 50, 7117–7131. PMID 21770428 Johnson B.B., Moe P.C., Wang D., Rossi K., Trigatti B.L., & Heuck A.P. (2012) Modifications in Perfringolysin O domain 4 alter the threshold of cholesterol concentration required for binding. Biochemistry, 51, 3373-3382. PMID 21770428 Pocognoni C.A., De Blas G.A., Heuck A.P., Belmonte S.A., Mayorga L.S. (2013) Perfringolysin O as a Useful Tool to Study Human Sperm Physiology Fertility and Sterility 99, 99-106. PMID: 22999796 Fahie M., Romano F.B., Chisholm C., Heuck A.P., Zbinden M., Chen M. (2013) A non-classical assembly pathway of Escherichia coli pore forming toxin cytolysin A. Journal of Biological Chemistry 288(43):31042-51. PMID: 24019520. Raghava, S.; Giorda, K. M.; Romano, F. B, Heuck A.P. Hebert D. (2013) SV40 Late Protein VP4 Forms Toroidal Pores To Disrupt Membranes for Viral Release. Biochemistry 52, 3939-3948. PMID: 23651212 Divakaruni AS, Wiley SE, Rogers GW, Andreyev AY, Petrosyan S, Loviscach M, Wall EA, Yadava N, Heuck AP, Ferrick DA, Henry RR, McDonald WG, Colca JR, Simon MI, Ciaraldi TP, Murphy AN. (2013) Thiazolidinediones are acute, specific inhibitors of the mitochondrial pyruvate carrier. Proceedings of the National Academy of Sciences (USA) 110, 5422-5427. PMID: 23513224 Other publications Book chapters Heuck A.P., Johnson A.E. (2005) “Membrane Recognition and Pore Formation by Bacterial Pore-forming Toxins”. In Protein-lipid interactions. L. Tamm Editor. Wiley-VHC. Heuck A. P., Moe, P. C., Johnson B.B. (2010) “The cholesterol-dependent cytolysins family of Gram-positive bacterial toxins” in Cholesterol-binding and Cholesterol Transport Proteins: Structure and Function in Health and Disease. Subcellular Biochemistry Series, Vol. 51, Chapter 20, 551-577. Edited by Harris R. Springer. PMID 20213558 Johnson B.B. and Heuck A.P. 2014 Perfringolysin structure and mechanism of pore formation as a paradigm for cholesterol-dependent cytolysins. In Pore Formation by MAPCF/CDC proteins- Agents of Defence, Attack and Invasion, Anderluh G. and Gilbert R. Editors. In press D. Research Support. Ongoing Research Projects NIH R01 GM097414 P.I.: Heuck A.P. 05/01/12-04/30/17 Molecular mechanism of Translocon Assembly into Cell Plasma Membranes Role: P.I. This project focuses on the molecular mechanism of translocon assembly into cell plasma membranes Completed Research Projects CEAR P.I.: Nagendra Yadava Center of Excellence in Apoptosis Research Translational Grant 07/01/11-11/30/12 Development of Perfringolysin O-based cell permeabilization kit for assay of mitochondrial function. Role: Co-PI This project focuses in the development of simple and reproducible assays for mitochondrial function, which can be commercialized as kits with the help of an industry partner (Seahorse Biosciences). P1FRG0000000101 P.I. : Alejandro P. Heuck University of Massachusetts Faculty Research Grant 12/01/10 – 11/30/11 Title: Structural determination of the Type III secretion translocon. Role: Principal investigator This project focuses on the in vitro reconstitution of the Pseudomonas aeruginosa Type III secretion translocon complex into membranes, and the analysis of the stoichiometry of membrane inserted translocons. AHA 0830136N P.I.: Alejandro P. Heuck American Heart Association Scientist Development Grant 01/01/08 – 12/31/11 Title: Development of novel probes for direct observation of cholesterol levels in mammalian cell membranes. Role: Principal Investigator This project focuses on the development of cellular probes to detect cholesterol levels on cell membranes
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