John Leszyk PhD
|Title||Research Associate Professor|
|Institution||University of Massachusetts Medical School|
|Department||Biochemistry and Molecular Pharmacology|
|Address||University of Massachusetts Medical School|
222 Maple Avenue
Shrewsbury MA 01545
Ph.D., Clarkson University, 1988
B.S., SUNY at Oswego, 1984
Proteomics and Mass Spectrometry
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.
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