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    John Leszyk PhD

    TitleResearch Associate Professor
    InstitutionUniversity of Massachusetts Medical School
    DepartmentBiochemistry and Molecular Pharmacology
    AddressUniversity of Massachusetts Medical School
    222 Maple Avenue
    Shrewsbury MA 01545
    Phone508-856-7533
        Overview 
        Narrative

        Academic Background

        Ph.D., Clarkson University, 1988
        B.S., SUNY at Oswego, 1984

        Proteomics and Mass Spectrometry

        Photo: John D. Leszyk I have been involved with analytical protein chemistry since my graduate work in the mid 1980’s where I used a first generation gas-phase Edman sequencer to determine amino acid sequences and chemical cross-linking sites of the subunits of the Troponin complex which are responsible for the Ca2+ regulation of muscle contraction. I then did a short post-doctoral study at the University Of Maryland School Of Medicine where I continued to pursue studies of the muscle regulatory subunits of the Troponin complex. In 1989 I came to the Worcester Foundation for Experimental Biology to set up their protein chemistry laboratory where I offered Edman sequencing, amino acid analysis and peptide synthesis services. Over time my approach to characterizing proteins evolved from a largely microchemical to a mass spectrometry based approach. The need for Edman sequencing diminished significantly due to the development of mass spectrometry based protein identification techniques which utilized both MS and tandem (MS/MS) data along with molecular weight search engine (MOWSE) software. In 1995 I acquired my first Matrix Assisted Laser Desorption/Ionization (MALDI) mass spectrometer. Since then I had developed a strong interest in the applications of MALDI time-of-flight (TOF) mass spectrometry in the area of protein characterization. In 1997 the Worcester Foundation merged with the University of Massachusetts Medical School where I became a research assistant professor in the department of Biochemistry and Molecular Pharmacology. My laboratory had continued as an independent self-supporting proteomics mass spectrometry core facility until 2010 when it was combined with two other proteomics facilities on campus to form an integrated Proteomics and Mass Spectrometry Facility. As Associate Director of the new facility I have been largely involved with the setup and implementation of various software pipelines for both the quantitative and qualitative aspects of proteomics analyses. I have also been involved with setup and optimization of the nanoflow LC/MS/MS experiment on the facility’s Thermo Velos and Q Exactive Orbitrap mass spectrometers. With the new instrumentation and software pipelines more complex proteomics experiments are now being carried out routinely by the facility. Such experiments inlcude characterization of post-translational modifications of complex samples, both label free and labeled quantitation experiments (iTRAQ, and SILAC), and the identification of chemical cross-links.

        Bibliographic 
        selected publications
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        1. Smith TC, Fridy PC, Li Y, Basil S, Arjun S, Friesen RM, Leszyk J, Chait BT, Rout MP, Luna EJ. Supervillin Binding to Myosin II and Synergism with Anillin Are Required for Cytokinesis. Mol Biol Cell. 2013 Oct 2.
          View in: PubMed
        2. Jones KA, Kim PD, Patel BB, Kelsen SG, Braverman A, Swinton DJ, Gafken PR, Jones LA, Lane WS, Neveu JM, Leung HC, Shaffer SA, Leszyk JD, Stanley BA, Fox TE, Stanley A, Hall MJ, Hampel H, South CD, de la Chapelle A, Burt RW, Jones DA, Kopelovich L, Yeung AT. Immunodepletion Plasma Proteomics by TripleTOF 5600 and Orbitrap Elite/LTQ-Orbitrap Velos/Q Exactive Mass Spectrometers. J Proteome Res. 2013 Sep 4.
          View in: PubMed
        3. Kiso WK, Selvaraj V, Nagashima J, Asano A, Brown JL, Schmitt DL, Leszyk J, Travis AJ, Pukazhenthi BS. Lactotransferrin in Asian Elephant (Elephas maximus) Seminal Plasma Correlates with Semen Quality. PLoS One. 2013; 8(8):e71033.
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        4. Singh G, Kucukural A, Cenik C, Leszyk JD, Shaffer SA, Weng Z, Moore MJ. The Cellular EJC Interactome Reveals Higher-Order mRNP Structure and an EJC-SR Protein Nexus. Cell. 2012 Oct 17.
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        5. Li Y, Powell DA, Shaffer SA, Rasko DA, Pelletier MR, Leszyk JD, Scott AJ, Masoudi A, Goodlett DR, Wang X, Raetz CR, Ernst RK. LPS remodeling is an evolved survival strategy for bacteria. Proc Natl Acad Sci U S A. 2012 May 29; 109(22):8716-21.
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        6. Koles K, Nunnari J, Korkut C, Barria R, Brewer C, Li Y, Leszyk J, Zhang B, Budnik V. Mechanism of Evi-exosome release at synaptic boutons. J Biol Chem. 2012 Mar 21.
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        7. Kruger AJ, Yang C, Tam SW, Hinerfeld D, Evans JE, Green KM, Leszyk J, Yang K, Guberski DL, Mordes JP, Greiner DL, Rossini AA, Bortell R. Haptoglobin as an early serum biomarker of virus-induced autoimmune type 1 diabetes in biobreeding diabetes resistant and LEW1.WR1 rats. Exp Biol Med (Maywood). 2010 Nov 1; 235(11):1328-37.
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        8. Leszyk JD. Evaluation of the new MALDI matrix 4-chloro-alpha-cyanocinnamic acid. J Biomol Tech. 2010 Jul; 21(2):81-91.
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        9. Romero R, Kusanovic JP, Gotsch F, Erez O, Vaisbuch E, Mazaki-Tovi S, Moser A, Tam S, Leszyk J, Master SR, Juhasz P, Pacora P, Ogge G, Gomez R, Yoon BH, Yeo L, Hassan SS, Rogers WT. Isobaric labeling and tandem mass spectrometry: a novel approach for profiling and quantifying proteins differentially expressed in amniotic fluid in preterm labor with and without intra-amniotic infection/inflammation. J Matern Fetal Neonatal Med. 2010 Apr; 23(4):261-80.
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        10. Zhou QL, Jiang ZY, Holik J, Chawla A, Hagan GN, Leszyk J, Czech MP. Akt substrate TBC1D1 regulates GLUT1 expression through the mTOR pathway in 3T3-L1 adipocytes. Biochem J. 2008 May 1; 411(3):647-55.
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        11. Davis MA, Hinerfeld D, Joseph S, Hui YH, Huang NH, Leszyk J, Rutherford-Bethard J, Tam SW. Proteomic analysis of rat liver phosphoproteins after treatment with protein kinase inhibitor H89 (N-(2-[p-bromocinnamylamino-]ethyl)-5-isoquinolinesulfonamide). J Pharmacol Exp Ther. 2006 Aug; 318(2):589-95.
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        12. Pantua H, McGinnes LW, Leszyk J, Morrison TG. Characterization of an alternate form of Newcastle disease virus fusion protein. J Virol. 2005 Sep; 79(18):11660-70.
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        13. Pazour GJ, Agrin N, Leszyk J, Witman GB. Proteomic analysis of a eukaryotic cilium. J Cell Biol. 2005 Jul 4; 170(1):103-13.
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        14. Jiang ZY, Zhou QL, Holik J, Patel S, Leszyk J, Coleman K, Chouinard M, Czech MP. Identification of WNK1 as a substrate of Akt/protein kinase B and a negative regulator of insulin-stimulated mitogenesis in 3T3-L1 cells. J Biol Chem. 2005 Jun 3; 280(22):21622-8.
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        15. Wilson-Fritch L, Nicoloro S, Chouinard M, Lazar MA, Chui PC, Leszyk J, Straubhaar J, Czech MP, Corvera S. Mitochondrial remodeling in adipose tissue associated with obesity and treatment with rosiglitazone. J Clin Invest. 2004 Nov; 114(9):1281-9.
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        16. Blüher M, Wilson-Fritch L, Leszyk J, Laustsen PG, Corvera S, Kahn CR. Role of insulin action and cell size on protein expression patterns in adipocytes. J Biol Chem. 2004 Jul 23; 279(30):31902-9.
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        17. Guilherme A, Soriano NA, Bose S, Holik J, Bose A, Pomerleau DP, Furcinitti P, Leszyk J, Corvera S, Czech MP. EHD2 and the novel EH domain binding protein EHBP1 couple endocytosis to the actin cytoskeleton. J Biol Chem. 2004 Mar 12; 279(11):10593-605.
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        18. Semiz S, Park JG, Nicoloro SM, Furcinitti P, Zhang C, Chawla A, Leszyk J, Czech MP. Conventional kinesin KIF5B mediates insulin-stimulated GLUT4 movements on microtubules. EMBO J. 2003 May 15; 22(10):2387-99.
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        19. Wilson-Fritch L, Burkart A, Bell G, Mendelson K, Leszyk J, Nicoloro S, Czech M, Corvera S. Mitochondrial biogenesis and remodeling during adipogenesis and in response to the insulin sensitizer rosiglitazone. Mol Cell Biol. 2003 Feb; 23(3):1085-94.
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        20. Nebl T, Pestonjamasp KN, Leszyk JD, Crowley JL, Oh SW, Luna EJ. Proteomic analysis of a detergent-resistant membrane skeleton from neutrophil plasma membranes. J Biol Chem. 2002 Nov 8; 277(45):43399-409.
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        21. Luo Y, Leszyk J, Li B, Li Z, Gergely J, Tao T. Troponin-I interacts with the Met47 region of skeletal muscle actin. Implications for the mechanism of thin filament regulation by calcium. J Mol Biol. 2002 Feb 22; 316(3):429-34.
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        22. Park JG, Bose A, Leszyk J, Czech MP. PYK2 as a mediator of endothelin-1/G alpha 11 signaling to GLUT4 glucose transporters. J Biol Chem. 2001 Dec 21; 276(51):47751-4.
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        23. Luo Y, Leszyk J, Li B, Gergely J, Tao T. Proximity relationships between residue 6 of troponin I and residues in troponin C: further evidence for extended conformation of troponin C in the troponin complex. Biochemistry. 2000 Dec 19; 39(50):15306-15.
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        24. Guilherme A, Emoto M, Buxton JM, Bose S, Sabini R, Theurkauf WE, Leszyk J, Czech MP. Perinuclear localization and insulin responsiveness of GLUT4 requires cytoskeletal integrity in 3T3-L1 adipocytes. J Biol Chem. 2000 Dec 8; 275(49):38151-9.
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        25. Luo Y, Leszyk J, Qian Y, Gergely J, Tao T. Residues 48 and 82 at the N-terminal hydrophobic pocket of rabbit skeletal muscle troponin-C photo-cross-link to Met121 of troponin-I. Biochemistry. 1999 May 18; 38(20):6678-88.
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        26. Choi JY, van Wijnen AJ, Aslam F, Leszyk JD, Stein JL, Stein GS, Lian JB, Penman S. Developmental association of the beta-galactoside-binding protein galectin-1 with the nuclear matrix of rat calvarial osteoblasts. J Cell Sci. 1998 Oct; 111 ( Pt 20):3035-43.
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        27. San Agustin JT, Leszyk JD, Nuwaysir LM, Witman GB. The catalytic subunit of the cAMP-dependent protein kinase of ovine sperm flagella has a unique amino-terminal sequence. J Biol Chem. 1998 Sep 18; 273(38):24874-83.
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        28. Leszyk J, Tao T, Nuwaysir LM, Gergely J. Identification of the photocrosslinking sites in troponin-I with 4-maleimidobenzophenone labelled mutant troponin-Cs having single cysteines at positions 158 and 21. J Muscle Res Cell Motil. 1998 Jun; 19(5):479-90.
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        29. Deyst KA, Bowe MA, Leszyk JD, Fallon JR. The alpha-dystroglycan-beta-dystroglycan complex. Membrane organization and relationship to an agrin receptor. J Biol Chem. 1995 Oct 27; 270(43):25956-9.
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        30. Cao QP, Pitt S, Leszyk J, Baril EF. DNA-dependent ATPase from HeLa cells is related to human Ku autoantigen. Biochemistry. 1994 Jul 19; 33(28):8548-57.
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        31. Bowe MA, Deyst KA, Leszyk JD, Fallon JR. Identification and purification of an agrin receptor from Torpedo postsynaptic membranes: a heteromeric complex related to the dystroglycans. Neuron. 1994 May; 12(5):1173-80.
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        32. Vissavajjhala P, Leszyk JD, Lin-Goerke J, Ross AH. Structural domains of the extracellular domain of human nerve growth factor receptor detected by partial proteolysis. Arch Biochem Biophys. 1992 Apr; 294(1):244-52.
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        33. Leszyk J, Grabarek Z, Gergely J, Collins JH. Characterization of zero-length cross-links between rabbit skeletal muscle troponin C and troponin I: evidence for direct interaction between the inhibitory region of troponin I and the NH2-terminal, regulatory domain of troponin C. Biochemistry. 1990 Jan 9; 29(1):299-304.
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        34. Leszyk J, Mornet D, Audemard E, Collins JH. Caldesmon structure and function: sequence analysis of a 35 kilodalton actin- and calmodulin-binding fragment from the C-terminus of the turkey gizzard protein. Biochem Biophys Res Commun. 1989 May 15; 160(3):1371-8.
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        35. Leszyk J, Mornet D, Audemard E, Collins JH. Amino acid sequence of a 15 kilodalton actin-binding fragment of turkey gizzard caldesmon: similarity with dystrophin, tropomyosin and the tropomyosin-binding region of troponin T. Biochem Biophys Res Commun. 1989 Apr 14; 160(1):210-6.
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        36. Leszyk J, Collins JH, Leavis PC, Tao T. Cross-linking of rabbit skeletal muscle troponin subunits: labeling of cysteine-98 of troponin C with 4-maleimidobenzophenone and analysis of products formed in the binary complex with troponin T and the ternary complex with troponins I and T. Biochemistry. 1988 Sep 6; 27(18):6983-7.
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        37. Leszyk J, Collins JH, Leavis PC, Tao T. Cross-linking of rabbit skeletal muscle troponin with the photoactive reagent 4-maleimidobenzophenone: identification of residues in troponin I that are close to cysteine-98 of troponin C. Biochemistry. 1987 Nov 3; 26(22):7042-7.
          View in: PubMed
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