Assistant Engineering

Jonathan P. B. Lees, Ph.D.

** ******* ****, *********, ** 02420 • 781-***-**** • *****@***.***

Objective

I am seeking a research position where I can apply my expertise in protein

purification, structural analysis, and engineering to address human health problems.

Education

Ph.D. in Biology, Johns Hopkins University, Baltimore, MD (2012)

B.A. in Chemistry, concentration in Biochemistry, Colby College, Waterville, ME (2005)

Highlighted Expertise and Skills

• Protein engineering by site-directed mutagenesis of twenty-two variants of the

mitochondrial fission protein Fis1

• Protein expression and purification of native and affinity tagged proteins

• Protein characterization using multiple spectroscopy techniques

• Proven leadership as a mentor of three undergraduates in the Hill lab (2008-2012)

and as a lab teaching assistant for Biochemistry and Cellular Biology courses (2006)

Research Experience

Mitochondrial fission protein dimerizes which has implications for its function (2005-2012)

Johns Hopkins University with R. Blake Hill, Ph.D.

• Discovered that the mitochondrial protein Fis1 homodimerizes.

• Determined the dimer topology using spectroscopic techniques.

• Engineered variants using site-directed mutagenesis that disrupt or enhance Fis1

dimerization.

• Applied a kinetic model to describe the dimerization of Fis1.

• Found that yeast cannot respire with the Fis1 mutations suggesting that an

interconversion between monomer and dimer is necessary for function.

The molecular basis of the proposed interaction of two small molecules (2004-2005)

Colby College with Thomas Shattuck, Ph.D.

• Showed that nizatidine and pyridoxine do not interact using isothermal titration

calorimetry contrary to a previous computational prediction.

The composition and packing of myelin (2002-2004

Boston College with Daniel Kirschner, Ph.D. Summers)

• Sequenced the signal sequence of the frog myelin protein zero.

• Found a carbohydrate moiety on the zebrafish myelin protein zero.

• Created a human myelin protein zero variant that causes a known packing defect and

optimized purification of the variant.

Publications

Lees JP, Manlandro CM, Picton L, Tan AE, Casares S, Flanagan JM, Fleming KG, Hill RB.

2012. A designed point mutant in Fis1 disrupts dimerization and mitochondrial fission. J

Mol Biol. 423: 143-158. Featured cover article.

Tooley JE, Khangulov V, Lees JP, Schlessman J, Bewley C, Heroux A, Bosch J, Hill RB.

2011. The 1.75 Å resolution structure of fission protein Fis1 from Saccharomyces

cerevisiae reveals elusive interactions of the autoinhibitory domain. Acta Crystallogr Sect

F Struct Biol Cryst Commun. 67: 1310-1315.

Luo X, Cerullo J, Dawli T, Priest C, Haddadin Z, Kim A, Inouye H, Suffoletto BP, Avila RL,

Lees JP, Sharma D, Xie B, Costello CE, Kirschner DA. 2008. Peripheral myelin of

Xenopus laevis: Role of electrostatic and hydrophobic interactions in membrane

compaction. J Struct Biol. 162: 170-83.

Avila RL, Tevlin BR, Lees JP, Inouye H, Kirschner DA. 2007. Myelin structure and

composition in zebrafish. Neurochem Res. 32: 197-209.

Posters Presented

The 24th Symposium of the Protein Society, San Diego, CA. The Dimerization of the (2010)

Cytosolic Domain of Fis1 May Regulate Mitochondrial Fission in Yeast.

The 21st Symposium of the Protein Society, Boston, MA. A Model for Fis1 (2007)

Regulation of Mitochondrial Dynamics.

Additional Expertise and Skills

• Protein Purification: affinity chromatography with immobilized metal ion affinity,

amylose, and glutathione columns, ion-exchange chromatography, and size exclusion

chromatography using the ÄKTAprime plus, the ÄKTA FPLC (automated

chromatography system), and the Agilent liquid chromatography HPLC systems.

• Molecular Biology: PCR, Western blot, and native gel electrophoresis.

• Spectroscopy: steady-state fluorometry, circular dichroism (CD), multi-angle light

scattering (MALS), matrix-assisted laser desorption/ionization time of flight mass

spectrometry (MALDI-TOF), and nuclear magnetic resonance (NMR) spectroscopy.

• Calorimetry: differential scanning calorimetry (DSC), isothermal scanning calorimetry

(ITC), and pressure perturbation calorimetry (PPC).

• Cytological Techniques: in yeast overexpressing native and tagged proteins, co-

immunoprecipitation, crosslinking, and mitochondrial imaging by fluorescence

microscopy.

• Software: Microsoft Office (Excel, Word, and PowerPoint) and the model fitting software

program IgorPro.

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