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Protein Research Scientist

Location:
Madison, WI
Posted:
November 14, 2014

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Resume:

**** ********* **** *** *

Madison, W isconsin ***11

Ann L. Menefee **********@*****.***

608-***-****

PROTEIN RESEARCH SCIENTIST

Highly motivated to design, perform, and analyze innovative protein purification solutions for complex and routine projects.

Performance Profile

• Over 10 years of experience directing concurrent projects that included optimizing protein purification and

characterizing the biochemical and biophysical properties of proteins.

• Five years of experience cloning and expressing proteins in prokaryotic and eukaryotic systems.

• Independent and dedicated analyst determined to achieve accurate, timely results whether working alone or as a

member of a collaborative team .

• Quick learner and creative problem-solver adept at developing new techniques to meet research goals.

• Strong communicator with proven written publication and oral presentation record.

Core Competencies

Protein Purification (Ӓkta FPLC)

Project Planning & Experimental Design

Protein Characterization (Activity & Binding Assays) Analytical Interpretation of Results

Molecular Biology (Cloning & Protein Expression) Mentoring, Training, & Support

Technical Writing & Oral Presentations Method Development

Laboratory Management Progressive Skill Improvement

PROFESSIONAL EXPERIENCE

CACTUS COMMUNICATIONS, Madison, WI • 2014 – Present

Freelance Editor

• Review grammar and spelling of scientific manuscripts for publication in the fields of Biochemistry, Molecular

Biology, and Genetics for an international editing service provider.

• Suggest changes in copy to enhance readability of documents and provide constructive feedback for future edits.

• Manage multiple edits with strict adherence to deadline conformity.

EAST CAROLINA UNIVERSITY/ BRODY SCHOOL OF MEDICINE, Greenville, NC • 2012 – 2013

Postdoctoral Scholar / Lab Manager

• Established a protein chemistry laboratory within 3 months with minimal supervision, including development and

maintenance of the laboratory procedures and protocols manual, safety guidelines, and equipment usage.

• Improved expression and enhanced purity of human transglutaminase II; characterized kinetic properties with

allosteric effectors.

• Generated and purified peptides to resolve complications of full-length substrate proteins (e.g. Tau).

• Designed and constructed a large-scale culture vessel to support 20 L bacterial cultures and to improve and

increase soluble protein expression.

• Supervised 5 technicians and graduate students in protein expression, purification, and assay development.

MARQUETTE UNIVERSITY, Milwaukee, W I • 2009 – 2012

Postdoctoral Fellow / Lab Manager

• Executed various cloning and protein engineering strategies to optimize expression, purification, and

crystallization, including the following: yeast and prokaryotic urea amidolyase, human and R. etli pyruvate

carboxylase, streptavidin, and biotin protein ligase.

• Generated a robust biotin auxotroph protein expression system to enable in vivo ligation of biotin analogs.

Ann L. Menefee **********@*****.*** PAGE 2

• Optimized coupled enzyme assays for mechanistic study with site-directed mutants of pyruvate carboxylase.

• Developed SOP for protein X-ray diffractometer and provided training and maintenance.

• Trained a lab of 5 students (graduate and undergraduate) in proper laboratory procedures; troubleshooting

cloning, protein purification, assay development, and crystallography techniques.

UNIVERSITY OF WISCONSIN / ENZYME INSTITUTE, Madison, W I • 2004 – 2009

Graduate Research Assistant

• Engineered three functional enzyme constructs to solve instability and solubility issues of ethanolamine ammonia

lyase for protein crystallography.

• Revealed mechanism of inactivation with substrates and analogs in the enzyme ethanolamine ammonia lyase.

• Delivered expertise in the generation and separation of hybrid protein dimers of enolase in a collaborative

mechanistic study.

• Purified eleven recombinant and native proteins (e.g. enolase, pyruvate: ferredoxin oxidoreductase, ferredoxin,

and pyruvate carboxylase) to homogeneity through size exclusion, hydrophobic, affinity, and ion exchange

chromatography.

• Cultivated a cooperative rapport with other labs by providing advice, training, and troubleshooting.

TEXAS A&M UNIVERSITY, College Station, TX • 2004

Research Technician

• Identified a 5-residue pathway for allosteric communication through analysis of the kinetic properties of mutant

proteins of phosphofructokinase.

• Devised protein purification schemes for 10 mutant proteins of phosphofructokinase.

• Contributed to the foundation and design of a Ph.D. thesis project.

FORMAL EDUCATION

Doctor of Philosophy in Biochemistry, 2009

University of Wisconsin – Madison, WI

Bachelor of Science in Biochemistry & Genetics, 2003

Texas A&M University – College Station, TX

SELECTED PUBLICATIONS

Menefee AL, Zeczycki TN. Nearly 50 years in the making: defining the catalytic mechanism of the multifunctional

enzyme, pyruvate carboxylase. FEBS J. 2014 Mar; 281(5):1333-54.

Zeczycki TN, Menefee AL, Jitrapakdee S, Wallace JC, Attwood PV, St Maurice M, Cleland WW. Activation and

inhibition of pyruvate carboxylase from Rhizobium etli. Biochemistry. 2011 Nov 15;50(45):9694-707.

Lietzan AD, Menefee AL, Zeczycki TN, Kumar S, Attwood PV, Wallace JC, Cleland WW, St Maurice M. Interaction

between the biotin carboxyl carrier domain and the biotin carboxylase domain in pyruvate carboxylase from

Rhizobium etli. Biochemistry. 2011 Nov 15; 50(45):9708-23.

Zeczycki TN, Menefee AL, Adina-Zada A, Jitrapakdee S, Surinya KH, Wallace JC, Attwood PV, St Maurice M,

Cleland WW. Novel insights into the biotin carboxylase domain reactions of pyruvate carboxylase from Rhizobium etli.

Biochemistry. 2011 Nov 15;50(45):9724-37.

Sims PA, Menefee AL, Larsen TM, Mansoorabadi SO, Reed GH. Structure and catalytic properties of an engineered

heterodimer of enolase composed of one active and one inactive subunit. J Mol Biol. 2006 Jan 20;355(3):422 -31

Paricharttanakul NM, Ye S, Menefee AL, Javid-Majd F, Sacchettini JC, Reinhart GD. Kinetic and structural

characterization of phosphofructokinase from Lactobacillus bulgaricus. Biochemistry. 2005 Nov 22;44(46):15280-6.



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