Ashlee N. Higdon. (205)

Transcription

1 Ashlee N. Higdon (205) Education PhD, Molecular and Cellular Pathology Program September 2011 University of Alabama at Birmingham Dissertation: Protein Modifications and the Response to Oxidized Lipids in Cardiovascular Cells Mentor: Victor Darley-Usmar, PhD Bachelors of Science in Chemical Engineering, Cum Laude May 2003 Biotechnology Emphasis University of Alabama in Huntsville Research Experience Postdoctoral Fellow 2013-present LSU-HSC, Laboratory of Dr. Imran Mungrue Study important biological functions and the regulation of the ER stress protein, CHAC1. Graduate student UAB, Laboratory of Dr. Victor Darley-Usmar Protein Modifications and the Response to Oxidized Lipids in Cardiovascular Cells Determine how the proteome is modified by endogenous lipid peroxidation using tagged lipids. Elucidate the effects of heme induced lipid peroxidation on mitochondrial function, autophagy and cell death. Examine the role of reactive lipid species on coordinating cellular processes such as upregulation of antioxidants and response to stress.

2 Research Assistant January 2005-April 2005 UAB, Department of Microbiology Used homologous recombination techniques using a suicide vector and conditionally replicating plasmid to delete the gene encoding the protein mpos, a hypothetical porin expressed in Mycobacterium Smegmatis, in the strain SMR5 (wild type M. Smegmatis). Designed primers for Southern Blot, PCR, and sequencing Purified chromosomal DNA from M. Smegmatis for PCR and Southern Blot Research Chemist I November 2003-January 2005 Biological & Physical Space Research Lab UAH, Marshall Space Flight Center Designed clones for overexpression of recombinant proteins. Pyruvate Dehydrogenase complex proteins were isolated and various protocols used for crystallization. Constructed and maintained a database with clone information, such as DNA sequences, primer design, protein domains, and hyperlinks to more information. Created protocols for the cell culture of myosarcoma and lung fibroblast cells obtained from ATCC, for use in radiation studies. Developed protocols for the studying the expression profiles of specific proteins. Used mrna obtained from myosarcoma cells and produced labeled cdna using RT-PCR. Performed microarray hybridization and analysis. Visiting Research Assistant June 2003-September 2003 Lab for Structural Biology University of Alabama in Huntsville Used genetic engineering techniques such as PCR, restriction digest, ligation, DNA purification, and transformation to express recombinant proteins. Both E.Coli and yeast expression systems were used. Clones were screened for expression levels of the protein Cathepsin K (human and the mouse homolog). Research Assistant June 2002-June 2003 Natural Products Drug Discovery Lab University of Alabama in Huntsville Took crude plant extract of the legume erythrina lanceolata from the Monte Verde cloud forest of Costa Rica, and performed several extraction and purification trials using organic solvent extractions, column chromatography, HPLC, and centrifugal TLC.

3 Awards and Funding Predoctoral Fellowship American Heart Association Modification of the macrophage proteome by oxidized lipids Young Investigator Award Nov Society of Free Radical Biology and Medicine Travel Awards: o Graduate Student Association Travel Grant 2008, 2010 o Pathology Travel Grant 2010 o Seahorse Biosciences Travel Award 2010 o Cayman Chemical Travel Grant Nov o Center for Free Radical Biology Travel Award Nov Professional Memberships and Honors Society for Free Radical Biology and Medicine 2005-present American Heart Association American Association for the Advancement of Science American Chemical Society Order of the Engineer 2003-present American Institute of Chemical Engineers EO Summer Internship Program, Mentor 2004 Marshall Space Flight Center, NASA Omega Chi Epsilon Chemical Engineering Honor Society Founding Member Beta Phi Chapter, 2001; Vice President,

4 College of Engineering Dean s List UAH Foundation Scholarship Recipient Poster Presentations Control of the Induction of Heme Oxygenase-1 by the Mitochondrially Targeted Antioxidant Mito-Tocopherol Pathology Graduate Student Research Day October 2009 Oxidized Arachidonic Acid Modifies Proteins in a Cyclooxygenase-2 Dependent Manner in Lipopolysaccharide-Stimulated Macrophage Cells Pathology Graduate Student Research Day October 2007 The Application of Novel Tagged Lipids to Determine the Mechanisms of Post- Translational Protein Modification by Oxidized Lipids Pathology Graduate Student Research Day October 2006 Expression of Proteins of the Pyruvate Dehydrogenase Complex Partnership for Biotechnology Bioretreat September 2004 Huntsville, AL Oral Presentations Effects of Protein Modification by Oxidized Lipids in Endothelial Cells Boston University June 2011 Boston, MA Hemin Induced Lipid Peroxidation Products Decrease Mitochondrial Function in Endothelial Cells Graduate Student Research Day February 2010

5 Hemin Induces Antioxidant Defenses Through the Formation of Oxidized Lipids: Importance of the Electrophile Responsive Proteome Graduate Student Research Day February 2009 Protein Modification and the Regulation of Antioxidants by Endogenous Products of Lipid Peroxidation Center for Free Radical Biology Seminar Series July 2008 Importance of Thiol Residues in Protein Modification by Electrophilic Lipids Graduate Student Research Day March 2006 Published Abstracts Bioenergetic Dysfunction in Endothelial Cells Precedes Cell Death due to Hemin: Implications for Hemolytic Anemias Society for Free Radical Biology and Medicine Conference November 2010 Orlando, FL Hemin Induces Antioxidant Defenses through the Formation of Oxidized Lipids: Importance of the Electrophile Responsive Proteome Society for Free Radical Biology and Medicine Conference November 2008 Indianapolis, IN Oxidized Arachidonic Acid Modifies Proteins in a Cyclooxygenase-2 Dependent Manner in Lipopolysaccharide-Stimulated Macrophage Cells Society for Free Radical Biology and Medicine November 2007 Washington, DC The Application of Novel Tagged Lipids to Determine the Mechanisms of Post- Translational Protein Modification by Oxidized Lipids *chosen for talk Society for Free Radical Biology and Medicine Conference November 2006 Denver, CO

6 Manuscripts Cummins TD, Higdon AN, Kramer PA, Chacko BK, Riggs DW, Salabei JK, Dell'italia LJ, Zhang J, Darley-Usmar VM, Hill BG. Utilization of fluorescent probes for the quantification and identification of subcellular proteomes and biological processes regulated by lipid peroxidation products. Free Radical Biology and Medicine. (Epub ahead of print); 2012 Higdon AN, Diers AR, Oh JY, Landar A, Darley-Usmar VM. Cell Signaling by Reactive Lipid Species: New Concepts and Molecular Mechanisms. Biochemical Journal. 442(3):453-64; 2012 Higdon AN, Landar A, Barnes S, Darley-Usmar VM. The Electrophile Responsive Proteome: Integrating Proteomics and Lipidomics with Cellular Function. Antioxidants and Redox Signaling. 17(11):1580-9; 2012 Higdon AN, Benavides GA, Chacko B, Ouyang X, Johnson MS, Landar A, Zhang J, Darley-Usmar VM. Hemin causes mitochondrial dysfunction in endothelial cells through promoting lipid peroxidation: The protective role of autophagy. AJP-Heart and Circulatory Physiology. 302(7):H ; 2012 *chosen for podcast Lee SH, Schneider C, Higdon AN, Darley-Usmar VM, Chung CY. Role of ipla(2) in the regulation of Src trafficking and microglia chemotaxis. Traffic. 12(7):878-89; 2011 Hill BG, Higdon AN, Dranka BP, Darley-Usmar VM. Regulation of vascular smooth muscle cell bioenergetics function by protein glutathiolation. Biochim Biophys Acta. 1797(2):285-95; 2010 Diers AR, Higdon AN, Ricard KC, Johnson MS, Agarwal A, Kalyanaraman B, Landar A, Darley-Usmar VM. Mitochondrial targeting of the electrophilic lipid 15-deoxy-Delta 12, 14-prostaglandin J2 increases apoptotic efficacy via redox cell signaling mechanisms. Biochem J. 426(1):31-41; 2010 Higdon AN, Dranka BP, Hill BG, Oh JY, Johnson MS, Landar A, Darley-Usmar VM. Methods for imaging and detecting modification of proteins by reactive lipid species. Free Radical Biology and Medicine. 47:201-12; 2009