Years Experience Project
Resume:
Michael D. Leipold
Toronto, ON M6G 2L5
************@*******.***
W: 416-***-**** H: 416-***-****
http://www.linkedin.com/in/mikeleipold
US citizen by birth.
.
.
EDUCATION AND WORK EXPERIENCE
Postdoctoral fellow Aug. 2007-present University of Toronto
Toronto, ON
Postdoctoral fellow Sept. 2003-Aug. 2007 University of Guelph
Guelph, ON
Ph.D. in Chemistry 1997-Sept. 2003 University of Utah
Salt Lake City, UT Graduate Coursework in Biological Chemistry
B.S. in Chemistry 1993-1997 Butler University
Indianapolis, IN ACS-certified degree
SKILLS
Enzyme purification - 9 years experience
Steady-state enzyme kinetics - 7 years experience
Pre-steady-state enzyme kinetics - 6 years experience, including the use of
rapid-quench
machinery (1 year)
Enzyme kinetics - inhibition assays, Ki determination, IC50 - 2 years
experience
FPLC protein purification - BioLogic system - 6 years experience
Liquid chromatography - ion exchange, affinity, gel filtration, hydrophobic
interaction - 8 years
experience
Molecular biology - cloning, protein overexpression, site-directed
mutagenesis - 8 years
experience
MALDI-TOF protein analysis - 2 years experience
Protein conjugation - 2 years experience
Microtiter plate assay - 2 years experience
In vivo complementation of bacterial systems - 2 years experience
Experimental handling of radioisotopes - 9 years experience
Electromobility shift assays - 2 years experience
Standard biochemical lab methods - PCR, gel electrophoresis, Western
blotting, UV/vis
spectroscopy - 9 years experience
Proficient in Adobe Photoshop and Illustrator, GraFit 4, Prism 4,
ImageQuant, Microsoft
Office.
Familiar with InsightII and DeepView.
PUBLICATIONS
Leipold, M.D., Herrera, I., Ornatsky, O., Baranov, V., Nitz, M., "ICP-MS-
Based Multiplex Profiling of Glycoproteins Using Lectins Conjugated to
Lanthanide-Chelating Polymers" J. Proteome Res., 2009, 8, 443-449
Leipold, M.D., Vinogradov, E., Whitfield, C., "Glycosyltransferases
Involved in Biosynthesis of the Outer Core Region of Escherichia coli
Lipopolysaccharides Exhibit Broader Substrate Specificities Than is
Predicted From Lipopolysaccharide Structures" J. Biol. Chem., 2007, 282,
Leipold, M.D., Kaniuk, N., Whitfield, C., "The C-terminal Domain of the
Escherichia coli WaaJ Glycosyltransferase is Important for Catalytic
Activity and Membrane Association" J. Biol. Chem., 2007, 282, 1257-1264
Leipold, M.D., Workman, H., Muller, J.G., Burrows, C.J., David, S.S.,
"Recognition and Removal of Oxidized Guanines in Duplex DNA by the Base
Excision Repair Enzymes hOGG1, yOGG1, and yOGG2" Biochemistry, 2003, 42,
Burrows, C.J., Muller, J.G., Kornyushnya, O., Luo, W., Duarte, V., Leipold,
M.D., David, S.S., "Structure and Potential Mutagenicity of New Hydantoin
Products from Guanosine and 8-Oxo-7,8-Dihydroguanosine Oxidation by
Transition Metals" Environ. Health Perspect., 2002, 110, 713-717.
Leipold, M.D., Muller, J.G., Burrows, C.J., David, S.S., "Removal of
Hydantoin Products of 8-Oxoguanine Oxidation by the Escherichia coli DNA
Repair Enzyme, FPG" Biochemistry, 2000, 39, 149**-*****.
PRESENTATIONS
Invited Talks
Leipold, M.D., Ornatsky, O.I., Baranov, V., Nitz, M.
"Single-Cell Resolution Profiling of Escherichia coli Strains Using Lectins
Conjugated to Lanthanide-Chelating Polymers"
Short talk at Candian Society for Chemistry 2010 Conference, Toronto, ON.
Leipold. M.D., Kaniuk, N.A., Whitfield, C.
"Role of the C-terminal Domain of E. coli WaaJ on Its Glycosyltransferase
Activity"
Short talk at FASEB Summer Research Conference - Microbial Polysaccharides
of Medical, Agricultural, and Industrial Importance, June 2006, Tucson, AZ.
Poster Presentations
Leipold, M.D., Herrera, I., Ornatsky, O., Baranov, V., Nitz, M., "ICP-MS-
Based Multiplex Profiling of Glycoproteins Using Lectins Conjugated to
Lanthanide-Chelating Polymers"
Poster presentation at Candian Society for Chemistry 2009 Conference,
Hamilton, ON.
Leipold. M.D., Kaniuk, N.A., Whitfield, C.
"Role of the C-terminal Domain of E. coli WaaJ on Its Glycosyltransferase
Activity"
Poster presentation at FASEB Summer Research Conference - Microbial
Polysaccharides of Medical, Agricultural, and Industrial Importance, June
2006, Tucson, AZ.
Leipold. M.D., Kaniuk, N.A., Whitfield, C.
"Investigation of the Effect of C-terminal Truncations on the Activity of
E. coli WaaJ"
Poster presentation at FASEB Summer Research Conference - Microbial
Polysaccharides of Medical, Agricultural, and Industrial Importance, June
2004, Tucson, AZ.
Leipold, M.D., Workman, H., Muller, J.G., Burrows, C.J., David, S.S.
"Removal of 8-Oxoguanine and Hydantoin Products by the DNA Repair Enzyme
Fpg" Poster presentation at the ACS National Meeting, Aug. 2001, Chicago,
IL.
DETAILED EXPERIENCE
Aug. 2007-present
University of Toronto Postdoctoral Fellow
Toronto, ON
Research Advisor: Professor Mark Nitz.
Project Overview: Conjugation of lectins with metal-binding polymers and
their use in single and multiplex format ICP-MS-based glycoprotein
profiling
* This has led to a first-author paper published in J. Proteome Res.
* Developed general protocol for the modification of lectins with metal-
binding polymers without loss of sugar-binding activity. This was in
extension of previous work from this lab involving labeling antibodies.
* Developed methods to reduce background signal by more than half relative
to initial assays.
* Developed and optimized a microtiter-plate-based assay to screen the
single and multiplex activity of the lectin-conjugates against commercial
glycoproteins.
Sept. 2003-Aug. 2007
University of Guelph Postdoctoral Fellow
Guelph, ON
Research Advisor: Professor Chris Whitfield.
Project Overview: Effect of C-terminal truncations on the activity of E.
coli glycosyltransferases involved in biosynthesis of outer core
lipopolysaccharide important for bacterial survival in the host
environment.
* This led to the publication of two first-author papers in J. Biol. Chem
* Used modeling and alignment with functionally homologous proteins to
determine sites of C-terminal protein truncation that would affect WaaJ
activity.
Used this information to create eight C-terminal truncation mutants in both
a high-expression and low-expression plasmid background to test the ability
of the C-terminal truncation mutants to complement an E. coli strain
deficient in waaJ as a function of in vivo protein concentration.
* Identified the subcellular localization of the E. coli
glycosyltransferases WaaR, WaaI, WaaO, WaaT, and each of the eight WaaJ
mutant proteins.
* Developed a protocol to overexpress and purify to homogeneity the
membrane-associated E. coli proteins WaaR, WaaI, WaaO, WaaT, and all WaaJ
mutant proteins.
* Isolated and purified lipopolysaccharide from two different E. coli
strains for use in in vitro reactions.
* Developed an in vitro assay to monitor activity of wild-type WaaT and
WaaJ C-terminal truncation mutant proteins by steady-state enzyme kinetics
using 14C-labeled substrates.
1997-Sept. 2003
University of Utah Graduate Research Assistant
Salt Lake City, UT
Research Advisor: Professor Sheila S. David
Project Overview: Recognition and repair of oxidized purines by the 8-
oxoguanine glycosylases Fpg, hOGG1, yOGG1, and yOGG2, enzymes involved in
the repair of oxidatively damaged DNA.
* This led to the publication of two first-author and one sixth-author
papers
* Collaborated with the Burrows lab at the University of Utah to study
secondary DNA base oxidation events and their effect on DNA structure and
recognition by DNA repair enzymes.
* Developed FPLC protocols to overexpress and purify the unmodified, wild-
type and two mutant E. coli Fpg proteins. This included the removal of a
co-purifying contaminant that degraded the in vitro substrate.
* Developed protocols to overexpress in E. coli and purify the histidine-
tagged eukaryotic glycosylases hOGG1, yOGG1, and yOGG2.
* Used alignment to detect amino acid residues conserved across homologous
proteins from twenty other bacteria, and created six mutants of Fpg.
* Used a combination of in vitro steady-state and pre-steady-state kinetics
in order to elucidate the differences in substrate recognition and cleavage
of three structurally distinct products of DNA oxidative damage by the DNA
repair glycosylases hOGG1, yOGG1, yOGG2, and wild-type and two mutants of
Fpg.
* Performed research for and wrote manuscripts of two first-author, peer-
reviewed articles, in addition to Ph.D. thesis.
* Directly oversaw the research training of one rotation and one graduate
student new to the David lab. Directly oversaw the research training of
two undergraduate students, including one for a period of two years.
PROFESSIONAL ACTIVITIES, AWARDS, AND MEMBERSHIPS
Honors, Awards
NIH Predoctoral Cross-Training Fellowship - July 1998 to June 1999
ACS Biological Division Graduate Research Travel Award - August 2001
William Epstein Graduate Fellowship in Chemistry - September 1997 - May
1998
University of Utah Graduate Research Supplemental Travel Award - August
2001
FASEB Summer Research Conference Travel Award - June 2006
Professional Affiliations
Sigma Xi (1997-present)
American Chemical Society (1997-present)
REFERENCES
Dr. Mark Nitz
University of Toronto
Department of Chemistry
80 St. George St.
Toronto, ON M5S 3H6
*****@****.********.**
Dr. Chris Whitfield
University of Guelph
Molecular and Cellular Biology
SC1 Rm: 1252
Guelph, ON N1G 2W1
********@********.**
519-***-**** x53361
Dr. Sheila David
University of California, Davis
Department of Chemistry
312 Chemistry Building
One Shields Avenue
Davis, CA 95616
*******@*******.***
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