Predoctoral Training Program

Annual Symposium

April 22, 2015

8:00 AM - 5:30 PM, Champions Club

Program Contacts

Director Melanie Simpson Rosowski Professor of Biochemistry N246 Beadle Center (402) 472-9309 msimpson2@unl.edu

Co-director Paul Black Bessey Professor and Chair of Biochemistry N200 Beadle Center (402) 472-3212 pblack2@unl.edu

Training Program Summary

The goal of the program is to develop outstanding new scientists who work in collaborative multi-disciplinary teams to research disease mechanisms using quantitative approaches that ultimately yield tangible strategies for prevention and therapy. Participants are highly qualified, motivated students with a strong interest in the underlying causes of human disease. The program provides a framework to encourage all students to assemble a broad knowledge base, actively seek research collaborations, produce an outstanding record of original published research, and develop presentation, proposal-writing, and leadership skills that position them for future excellence as independent researchers focused on mechanisms of disease progression.

Molecular Mechanisms of Disease Internal Advisory Committee

Prem Paul Vice Chancellor for Research and Economic Development

Ronnie Green NU Vice President Harlan Vice Chancellor, Institute of Agriculture and Natural Resources

Brian Larkins Associate Vice Chancellor for the Life Sciences

Laurie Bellows Associate Dean of Graduate Studies Director, McNair Program

Charles Wood Lewis Lehr Professor of Biological Sciences and Biochemistry Director, Center for Virology

Theresa Welbourne FirsTier Banks Distinguished Professor of Business Director, Center for Entrepreneurship President and CEO, eePulse, Inc.

The University of Nebraska – Lincoln is an equal opportunity educator and employer.

If you require special accommodations, please contact Melanie Simpson at msimpson2@unl.edu.

Molecular Mechanisms of Disease Mentoring Faculty

Jiri Adamec, Biochemistry Mitochondrial dysfunction biomarkers link to environmental stressors

Jim Alfano, Biological Sciences Suppression of innate immunity by an ADP-ribosyltransferase type III effector

Joseph Barycki, Biochemistry Structural insights into redox homeostasis

Don Becker, Biochemistry Role of proline in redox homeostasis and apoptosis

Andrew Benson, Food Science and Technology Genetic control over the gut microbiome composition

David Berkowitz, Chemistry Chemical biology and synthetic organic chemistry

Paul Black, Biochemistry Fatty acid transport in eukaryotes

Deb Brown, School of Biological Sciences Vaccine strategies that target cytolytic CD4 T cells to the lung

Nicole Buan, Biochemistry Contribution of methanogenic bacteria to gut function

Andrea Cupp, Animal Science Role of VEGF in testis morphogenesis

Concetta DiRusso, Biochemistry High throughput screens for fatty acid uptake inhibitors

Steve DiMagno, Chemistry Synthesis of novel fluorinated tumor imaging agents

Eric Dodds, Chemistry Chemical glycobiology

Liangcheng Du, Chemistry Discovering new anti-infective agents from Lysobacter

Jiantao Guo, Chemistry Protein tyrosine-O-sulfation, molecular interaction probes

David Hage, Chemistry Chromatographic automation of immunoassays

Ed Harris, Biochemistry Liver endothelial scavenger receptor function; systemic clearance of heparin

Oleh Khalimonchuk, Biochemistry Mitochondrial oxidative function and protein homeostasis in health and sickness

Sri Kidambi, Chem & Biomol Engineering Tissue engineering, stem cells, and liver reconstruction

Jaekwon Lee, Biochemistry Mechanistic insights in homeostatic copper acquisition and cellular metal detoxification

Angela Pannier, Biol Systems Engineering Non-viral gene delivery, synthetic extracellular matrices, protein-cell adhesion

Robert Powers, Chemistry NMR metabolomics

Amanda Ramer-Tait, Food Science and Technology Host-microbial interactions in inflammatory bowel disease

Wayne Reikhof, Biological Sciences Lipid distribution and storage mechanisms

Melanie Simpson, Biochemistry Steroid elimination, hyaluronan signaling and turnover in prostate cancer progression

Cliff Stains, Chemistry Novel fluorescent protein design and protein activity switches

James Takacs, Chemistry Catalytic asymmetric hydroboration and “green chemistry” synthesis

Bill Velander, Chem & Biomol Engineering cGMP recombinant factor IX for IV and oral hemophilia B therapy

Mark Wilson, Biochemistry Redox regulation of DJ-1 function

Jennifer Wood, Animal Sciences Steroid hormone sensing and signaling

Schedule of events

WEDNESDAY, April 22, 2015

8:00 AM TO 5:30 PM

University of Nebraska Alumni Champion’s Club

8:00-8:30 AM Poster set-up and continental breakfast

8:30-8:45 AM Morning session welcome, Melanie Simpson, Director

8:45-9:00 AM Morning session kickoff

Associate Vice Chancellor Steve Goddard

9:00-10:00 AM Morning plenary presentation

Celia Goulding, University of California - Irvine Structural insights into Mechanisms of Bacterial Survival

10:00-11:15 AM Student presentations I

Shelbi Christgen, Biochemistry Exploring the role of proline metabolism in Helicobacter pathogenicity

Christina Davis, Chemical and Biomolecular Engineering Stiffness induced reactive astrocytes: an in vitro model for studying astrogliosis Poster #4, Iryna Bohovych, Biochemistry

Metalloprotease Oma1 fine-tunes mitochondrial bioenergetic function and respiratory supercomplex stability

Poster #17, Sarah Romereim, Animal Science Granulosa Cell Cycle Regulation and Steroidogenesis in a High

Androstenedione Follicular Microenvironment

11:15-12:15 Session I poster viewing:

Molecular signaling and Oxidative stress

12:30-1:30 Lunch

1:30-2:30 PM Afternoon plenary presentation

John Denu, University of Wisconsin - Madison Newly Discovered Pathways that Regulate Metabolism and Healthy Aging

2:30-3:45 PM Student presentations II

Ryan Matsuda, Chemistry Overview of tools and techniques for characterization of the metabolomics effects of diabetes on serum proteins

Michelle Palmer, Biochemistry UDP-glucuronate partitioning in metabolic detoxification

Poster #30 Carlos Gomes Neto, Food Science & Technology Role of the resident microbiota in pathobiont-mediated susceptibility to intestinal inflammation

Poster #46 Alex Vogel, Biological Sciences Interferon regulatory factor 3 deficiency contributes to impaired memory T cell function in response to influenza infection

3:45-4:45 PM Session II poster viewing:

Disease microenvironment and Metabolic integrity

5:00 PM Award presentations

5:15-6:00 PM Poster take-down

PLENARY SPEAKER PROFILES

Celia W. Goulding received a B.Sc(Hons) in chemistry and mathematic and a Ph.D. in physical organic chemistry from King’s College, London. After leaving the UK for the US, she was a postdoctoral fellow with Professor Rowena Mathews at the University of Michigan studying metalloprotein enzymology. Moving further west, as research faculty at UCLA under the supervision of Professor David Eisenberg, Celia became interested in the mechanisms by which Mycobacterium tuberculosis, the bacterial pathogen which causes tuberculosis, is able to survive in human hostile environments, employing X-ray crystallography and other biophysical and biochemistry techniques. Celia eventually accepted an independent position at the University of California – Irvine,

where she and her group investigate the function and structure of metalloproteins in a recently discovered and unique heme uptake pathway in M. tuberculosis. She has more recently turned her attention to the structural characterization of components involved in bacterial contact-dependent growth inhibition systems.

John Denu received his B.S. degree from the University of Wisconsin–Madison and his Ph.D. from Texas A & M University, followed by a postdoctoral fellowship at the University of Michigan Medical School. He is currently Professor of Biomolecular Chemistry at the University of Wisconsin – Madison, and theme director of Epigenetics in the Wisconsin Institute for Discovery.

Dr. Denu’s research group has investigated the mechanisms and biological function of reversible protein modifications involved in modulating signal transduction, chromatin dynamics and metabolism. His group provided the first evidence that protein acetylation controls metabolic pathways, and that the physiological benefits of caloric

restriction are mediated in part by the sirtuin family of NAD+-dependent protein deacetylases. More recently, his group has demonstrated that dietary fatty acids can stimulate the activity of histone deacetylases, leading to the down regulation of genes involved in fat and carbohydrate metabolism.

Oral Presentation Abstracts

Morning Session

Trainee presentation #1

EXPLORING THE ROLE OF PROLINE METABOLISM IN HELICOBACTER PATHOGENICITY

Shelbi Christgen1, Amanda Ramer-Tait2, Don Becker1

1 Department of Biochemistry, University of Nebraska-Lincoln

2 Department of Food Science and Technology, University of Nebraska-Lincoln

Helicobacters have been associated with many diseases; including peptic ulcer disease, inflammatory bowel disease, hepatitis, and cancer. It is thought that the chronic inflammatory response experienced by the host in response to Helicobacter infection is the cause of subsequent host disease. One of the preferred respiratory substrates of Helicobacters is the amino acid proline. In some bacteria, including Helicobacter, proline is metabolized by an enzyme called proline utilization A (PutA). PutAs have been shown to utilize substrate channeling between two active site domains: a proline dehydrogenase (PRODH) domain and a pyrroline-5-carboxylate dehydrogenase (P5CDH) domain. The physiological importance of substrate channeling in any system is currently unknown. Previously, it has been shown that PutA is required for H. pylori infection and that disrupting the putA gene in H. hepaticus leads to decreased host inflammation. Among genes required for H. pylori infection are the genes coding for the peptidase collagenase and the proline transporter PutP. Due to the high proline percentage of collagen and the high levels of extracellular proline in hosts infected with Helicobacter species, we hypothesize that collagenase is required for infection because it provides the pathogen with a steady source of proline. Furthermore, we propose the breakdown of collagen results in chronic host inflammation and later disease. We seek to explore the relationship between collagenase and proline metabolism in the murine pathogen Helicobacter bilis. We also seek to elucidate the role of P5C intermediate channeling in H. bilis pathogenicity.

Trainee presentation #2

STIFFNESS INDUCED REACTIVE ASTROCYTES: AN IN VITRO MODEL FOR STUDYING ASTROGLIOSIS

Christina Wilson and Srivatsan Kidambi

Department of Chemical and Biomolecular Engineering, University of Nebraska

Studies have shown that the brain, a soft tissue highly susceptible to injury, can increase in stiffness as a result of certain injury and disease. Astrocytes, the most abundant non-neuronal cell type in the brain, have been observed to respond to brain injury and disease by reverting to an activated phenotype resulting in astrogliosis which has many beneficial and harmful molecular characteristics toward restoration of healthy brain function. The activated astrocyte phenotype has been studied under numerous chemical cues. However the reactive phenotype resulting from mechanical cues, such as the change in tissue stiffness observed in the event of traumatic brain injury, tumor progression and stroke, is not well documented. Therefore the aim of this study is to utilize an in vitro model to study the molecular mechanisms initiating and sustaining reactive astrocyte phenotype resulting from an increase in surface mechanical stiffness. We observed a stiffness dependent change in morphology and intermediate filament protein expression characteristic of astrogliosis, as well as increase in actin stress fibers, intercellular reactive oxygen species and loss of astrocyte glutamate uptake. These observation suggest that this model will be useful in further molecular mechanistic studies of astrogliosis.

Poster #4 Talk

METALLOPROTEASE OMA1 FINE-TUNES MITOCHONDRIAL BIOENERGETIC FUNCTION AND RESPIRATORY SUPERCOMPLEX STABILITY

Iryna Bohovych1,2, Mario R. Fernandez4, Jennifer J. Rahn5, Krista D. Stackley5, Jennifer E. Bestman5, Annadurai Anandhan2,3, Rodrigo Franco2,3, Steven M. Claypool6, Robert E. Lewis4, Sherine S. L. Chan5 and Oleh Khalimonchuk1,2

1 Department of Biochemistry, 2Nebraska Redox Biology Center and 3School of Veterinary Medicine and Biomedical Sciences, University of Nebraska-Lincoln, Lincoln NE, 68588. 4 Eppley Institute for Research in Cancer, University of Nebraska Medical Center, Omaha, NE 68198. 5 Department of Drug Discovery and Biomedical Sciences, Medical University of South Carolina, Charleston, SC 29425. 6 Department of Physiology, Johns Hopkins School of Medicine, Baltimore, MD 21205.

Normal mitochondrial functioning is assured through a complex network of proteases and chaperons termed the intramitochondrial quality control (IMQC) system. Despite high evolutionary conservation and growing research interest, functions of several IMQC modules remain unclear. Exemplary, little is known about an ATP-independent IM metalloprotease Oma1, which is involved in maintenance of adequate stress response and control of mitochondrial dynamics in eukaryotes.

Here we show that loss of Oma1 activity in yeast model leads to alterations in mitochondrial membrane potential, unbalanced respiration, as well as progressive respiratory decline during aging. The latter may be linked to the loss of respiratory supercomplexes stability (RSCs). Overexpression of RSCs-stabilizing factor Rcf1 suppresses the defect, whereas inactivation of this protein in Oma1-deficient cells leads to further destabilization. In addition, strong genetic interactions between Oma1 and other RSCs-stabilizing factors like ATP/ADP carrier Aac2 are observed in the yeast model.

Similarly, impaired supercomplex stability is associated with Oma1 inactivation in mouse embryonic fibroblasts (MEFs). Moreover, the Oma1-deficient MEFs are unable to maintain proper respiration under conditions, which require maximal mitochondrial capacity. Similar defects are seen upon Oma1 depletion in zebrafish vertebrate model, wherein loss of Oma1 is associated with developmental defects specific for impaired mitochondrial bioenergetics.

Taken together, our results suggest a novel role of Oma1 in fine-tuning of respiratory function and bioenergetics plasticity in eukaryotes.

 

Poster #17 Talk GRANULOSA CELL CYCLE REGULATION AND STEROIDOGENESIS IN A HIGH ANDROSTENEDIONE FOLLICULAR MICROENVIRONMENT

Sarah M. Romereim1, Adam F. Summers2, Bill E. Pohlmeier1, Renee M. McFee1, Renata Spuri-Gomes1, Scott G. Kurz1, Anthony K. McNeel3, Robert A. Cushman3, John S. Davis4, Jennifer R. Wood1, Andrea S. Cupp1

1University of Nebraska–Lincoln; 2New Mexico State University, 3USDA, ARS, U.S. Meat Animal Research Center, 4University of Nebraska Medical Center

Anovulatory infertility affects up to 40% of infertile women. In fact, anovulation may often go undetected considering that 8-13% of normally menstruating women have sporadic anovulation. While many of the more severe types of infertility disorders are currently being investigated, there has not yet been an animal model described with which to study the mechanisms of sporadic anovulation. In order to fill this model system gap, our lab has identified a naturally-occurring bovine model which includes a subpopulation of cows (~10%) with a 17% reduction in calving rate as well as endocrine and follicular development profiles resembling those of women with chronic or sporadic anovulation.

These cows with sporadic anovulation characteristics (designated SPOV) have high intrafollicular androstenedione (30-fold higher) in the dominant follicle compared to ovulatory cows (OV) but lack a corresponding increase in estradiol (2-4 fold). Affymetrix Bovine GeneChip 1.0 ST microarray analyses comparing ovarian somatic cells from SPOV and OV cows indicated that granulosa cells were undergoing cell cycle arrest. Using Ingenuity Pathway Analysis, we found that granulosa cells from the High A4 population exhibit a strong inhibition of the cell cycle, especially G1/S checkpoint proteins and regulators of chromosome alignment and segregation. Further experiments suggested that SPOV follicles do not maintain a sufficient population of granulosa cells to efficiently convert androgens to estrogen. This cow population with excess intrafollicular androgen and sporadic anovulation is an excellent model to study mechanisms involved in female anovulatory infertility and ultimately develop therapeutics and procedures to enhance reproductive success in women.

Oral Presentation Abstracts

Afternoon Session

Trainee presentation #3 OVERVIEW OF TOOLS AND TECHNIQUES FOR CHARACTERIZATION OF THE METABOLOMIC EFFECTS OF DIABETES ON SERUM PROTEINS

Ryan Matsuda, Venkata Kolli, Zhao Li, Megan Woods, Xiwei, Zheng, K.S. Hoy, Jeanethe Anguizola, Omar Barnaby, Eric D. Dodds, David S. Hage

Department of Chemistry, University of Nebraska, Lincoln, NE

The purpose of this study was to employ new analytical tools and techniques based on high-performance affinity chromatography and mass spectrometry to characterize the metabolomic effects of diabetes on the structure and function of human serum albumin (HSA). Diabetes is a disease that results in elevated levels of glucose in the bloodstream. As a result, diabetes also leads to the non-enzymatic glycation of serum proteins such as HSA. This study investigated the use of high-performance affinity chromatography to examine the binding of several sulfonylurea drugs to HSA at various levels of glycation. The results indicated that the level of glycation can affect the binding of this protein to these drugs. Electrospray ionization mass spectrometry (ESI-MS) was used to provide high resolution analysis of the structure of HSA and glycated HSA. Further investigation of the structure of glycated HSA through the use of MS/MS techniques provided more detailed information about the location and types of glycation-related modifications that occurred on this protein. The information obtained from these methods should provide biomedical researchers with a better understanding of how metabolomic diseases such as diabetes can affect the structure and function of serum proteins.

Trainee presentation #4

UDP-GLUCURONATE PARTITIONING IN METABOLIC DETOXIFICATION

Michelle E. Palmer, Jonathan E. Markham, Joseph J. Barycki, and Melanie A. Simpson

Department of Biochemistry, University of Nebraska, Lincoln, NE

UDP-glucuronate, a vital metabolite in prostate tumor cells, is generated by NAD+-dependent oxidation of UDP-glucose, catalyzed by UDP-glucose dehydrogenase (UGDH). UDP-glucuronate can be directed to hyaluronan synthesis at the plasma membrane, to proteoglycan production in the Golgi, or to the cellular detoxifying enzymes, UDP-glucuronosyltransferases, in the ER. In the prostate, cytosolic UDP-glucuronate levels can dictate the rate of glucuronidation of dihydrotestosterone (DHT) and xenobiotics to inactivate and eliminate excess steroids and lipophilic compounds in a highly regulated clearance mechanism. Despite the clearly observable readouts of each fate, the mechanisms by which UDP-glucuronate is directed into one or more pathways are not known. Perturbing these pathways with metabolic, chemical, and genetic approaches, followed by use of LCMS and cellular assays elucidates how the cells regulate and direct UDP-glucuronate into each metabolic fate. Having achieved 80% knockdown of downstream enzyme UDP-xylose synthase (UXS1) in prostate cancer PC3 cells, we optimized an LCMS approach to measure the UDP-sugar metabolites. Treatment with the chemical WP631 dihydrochloride has also achieved 40% inhibition of both UGDH and UXS1. Quantifying UDP-sugar levels across a variety of prostate cancer cell lines reveals suppressed glucuronidation in steroid sensitive lines and increased glucuronidation and hyaluronan production in developed prostate cancer lines. Evaluating metabolites and products of the UDP-sugar pathway is beginning to reveal the mechanisms by which cells partition UDP-glucuronate into a respective fate.

Poster #30 Talk ROLE OF THE RESIDENT MICROBIOTA IN PATHOBIONT-MEDIATED SUSCEPTIBILITY TO INTESTINAL INFLAMMATION

J. C. Gomes Neto1, H. Kittana1, L. B. Bindels1, R. Segura Munoz1, J. M. Hostetter2 and A. E. Ramer-Tait1

1Department of Food Science and Technology, University of Nebraska-Lincoln, Lincoln, NE, USA and 2Department of Veterinary Pathology, Iowa State University, Ames, IA, USA

Inflammatory bowel disease (IBD) behaves as multifactorial disease, resulting from interactions between environmental factors (i.e., gut microbiota) and host genetics. Recent studies have linked changes in microbiota composition not only with the onset of IBD but also with expansion of bacterial pathobionts. These opportunistic organisms, including enterohepatic Helicobacter species, do not cause disease in healthy individuals but can promote immune-mediated pathology during dysbiosis. How and why pathobionts selectively mediate disease in susceptible individuals is not known. Our aim is to define the mechanisms by which intestinal bacteria, including pathobionts, promote disease onset. To specifically assess the role of the resident microbiota in pathobiont-induced susceptibility to a mild inflammatory trigger, germ free (GF) and defined microbiota (altered Schaedler flora or ASF) C3H/HeN mice were colonized with Helicobacter bilis. An inflammatory trigger was then administered in the form of a sub-pathological dose of dextran sulfate sodium (DSS, 1.5%) not sufficient to elicit disease in the absence of the pathobiont. H. bilis-colonized ASF mice developed a more severe typhlocolitis following DSS treatment as compared to H. bilis-colonized GF mice. The presence of a microbiota also increased production of IL-1β, IL-33, IL-17A, IL-17F, TGF-β1 and IFN-γ and decreased IL-10 production in the cecum of mice colonized with H. bilis and treated with DSS. Moreover, ASF-bearing mice colonized with the Helicobacter pathobiont and exposed to the inflammatory trigger developed a population of CD4+ T cells that produced IL-17A, IL-17F, and IL-22 in response to specific members of the ASF community. Changes in the relative abundance of ASF members in luminal cecal contents did not explain the enhanced immunoreactivity to certain resident species. Together, these results indicate the resident microbiota exacerbates disease and fails to prevent immune deviation in the presence of both pathobiont and inflammatory trigger. Consequently, certain bacterial species become the target of an exacerbated Th17 immune response. These findings indicate an important contextual role of the microbiota in development of immune responses contributing to pathobiont-mediated disease susceptibility.

Poster #46 Talk INTERFERON REGULATORY FACTOR 3 DEFICIENCY CONTRIBUTES TO IMPAIRED MEMORY T CELL FUNCTION IN RESPONSE TO INFLUENZA INFECTION

Alexander J. Vogel1,2 Tyler C. Moore1,2 Thomas M. Petro3 and Deborah M. Brown1,2

1 School of Biological Sciences 2 Nebraska Center for Virology, University of Nebraska-Lincoln 3 Department of Oral Biology, University of Nebraska Medical Center The role of interferon regulatory factor 3 (IRF3) in initiating the type I interferon response in innate immune cells has been well studied in the context of viral infection. However, it remains unclear what impact IRF3 has in shaping and maintaining adaptive immune responses. To address this, IRF3 deficient (IRF3 KO) mice were infected with influenza A virus (IAV) and T cell responses measured. At day 9 post infection, the frequency of interferon gamma (IFN-γ) and Granzyme B (GrB) positive T cells were similar between IRF3 KO and wildtype mice. Despite similar primary responses, IRF3 KO mice exhibited significant deficits in recall memory T cell responses to IAV with reduced expression of IFN-γ and GrB. Since IRF3 has been implicated in the expression of cytokines that influence effector and memory T cells responses, cytokine supplementation studies were performed. Addition of IL-12 was the only cytokine tested that was able to rescue the IFN-γ deficiencies in IRF3 KO T cells. Interestingly, peptide specific GrB re-expression could not be restored by any cytokines tested. These results suggest that IRF3 plays a role in the maintenance of antigen specific memory T cell responses and that IRF3 deficiency has a substantial impact on the cytotoxic potential of memory T cells and recall responses. Together, this work provides further insight into the molecular regulation of adaptive immune responses to viral infections and may serve to inform future vaccine strategies that require the development of memory T cells.

Poster Presentations Morning Session:

Molecular signaling and Oxidative stress

1

COMBINING A CLOSTRIDIAL ENZYME EXHIBITING UNUSUAL ACTIVE SITE PLASTICITY WITH A REMARKABLY FACILE SIGMATROPIC REARRANGEMENT: RAPID, STERECONTROLLED ENTRY INTO DENSELY FUNCTIONALIZED FLUORINATED PHOSPHONATES FOR CHEMICAL BIOLOGY Gregory A. Applegate; Kaushik Panigrahi; David B. Berkowitz Department of Chemistry, University of Nebraska

2

VASCULAR ENDOTHELIAL GROWTH FACTOR A 165 (VEGFA165), ANGIOGENIC ISOFORM, PROMOTES WHILE VEGFA165b ANTAGONIZES VEGFA165 STIMULATED FOLLICULAR PROGRESSION IN BOVINE OVARIAN CORTICAL PIECES CULTURED FROM PRE-PUBERTAL HEIFERS Mohamed Ayoub1, Manjula Magamage3, Rebecca Vinton1, Robert A Cushman2, Anthony K. McNeel2, Harvey C. Freetly2, Jennifer R. Wood1, Andrea S. Cupp1 1 Department of Animal Science, University of Nebraska-Lincoln, Lincoln, Nebraska 2 USDA, ARS, U.S. Meat Animal Research Center, Clay Center, NE 68933 3 Department of Livestock Production, Sabaragamuwa University of Sri Lanka

3

PROFILING THE PHOSPHATOME WITH DIRECT ACTIVITY SENSORS Jon R. Beck1, Edward N. Harris2 and Cliff I. Stains1 1 Department of Chemistry, University of Nebraska 2 Department of Biochemistry, University of Nebraska

4

METALLOPROTEASE OMA1 FINE-TUNES MITOCHONDRIAL BIOENERGETIC FUNCTION AND RESPIRATORY SUPERCOMPLEX STABILITY Iryna Bohovych1,2, Mario R. Fernandez4, Jennifer J. Rahn5, Krista D. Stackley5, Jennifer E. Bestman5, Annadurai Anandhan2,3, Rodrigo Franco2,3, Steven M. Claypool6, Robert E. Lewis4, Sherine S. L. Chan5 and Oleh Khalimonchuk1,2 1 Department of Biochemistry, 2Nebraska Redox Biology Center and 3School of Veterinary Medicine and Biomedical Sciences, University of Nebraska-Lincoln; 4 Eppley Institute for Research in Cancer, University of Nebraska Medical Center; 5 Department of Drug Discovery and Biomedical Sciences, Medical University of South Carolina, Charleston, SC; 6 Department of Physiology, Johns Hopkins School of Medicine, Baltimore, MD.

5

MONITORING PROTEIN KINASE PERTURBATIONS DIRECTLY IN CELL CULTURE BY HIGH-THROUGHPUT CHELATION-ENHANCED FLUORESCENCE Garrett R. Casey1, Xinqi Zhou1, Cliff I. Stains1 1 Department of Chemistry, University of Nebraska, Lincoln, NE

6

DISCOVERY OF THE MEMBRANE BINDING DOMAIN IN THE PROLINE UTILIZATION A (PUTA) ENZYME Shelbi Christgen1,2, Weidong Zhu1,2, Bushra Bibi2, John Tanner3, Don Becker2 1 These authors contributed equally to this work

2 Department of Biochemistry, University of Nebraska-Lincoln 3 Departments of Biochemistry and Chemistry, University of Missouri-Columbia 7

AAA+ ATPASE AFG1 IS A NOVEL MEMBER OF THE MITOCHONDRIAL QUALITY CONTROL Edward Germany, Iryna Bohovych, Nataliya Zahayko and Oleh Khalimonchuk Department of Biochemistry and Redox Biology Center, University of Nebraska, Lincoln, NE 8

β-D-carbafructopyranosyl-1,2-diamine-derived salen emerges as a promising new chiral ligand from a miniaturized enzymatic screen (ISES) Kannan Karukurichi, Xiang Fei, Robert Swyka, Sylvain Broussy, Weijun Shen, Sangeeta Dey, Sandip Roy, David Berkowitz Department of Chemistry, University of Nebraska, Lincoln, Nebraska, United States 9

OPTIMIZATION OF A CHEMOSENSOR FOR RHO-ASSOCIATED PROTEIN KINASE AND ITS APPLICATION TO INHIBITOR SCREENING Maia I. Kelly1, Tyler J. Bechtel1,2, D. Rajasekhar Reddy1,3, Erome D. Hankore1, Jon R. Beck1, and Cliff I. Stains1 1 Department of Chemistry, University of Nebraska – Lincoln, Lincoln, Nebraska 68588 2 Department of Chemistry, Boston College, Chestnut Hill, Massachusetts 02467 3 Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine, St. Louis, Missouri 63110 10

EXPLORING THE IMPORTANCE OF DIMERIZATION FOR DJ-1 THROUGH ENGINEERED DOMAIN FUSIONS Jiusheng Lin, Sierra Hansen, Mark Wilson Department of Biochemistry, University of Nebraska-Lincoln 11

DEVELOPMENT AND DEPLOYMENT OF A NEW METHOXYVINYL CATION EQUIVALENT Christopher D. McCune, Matthew L. Beio, Jacob A. Friest, Sandeep Ginotra, David B. Berkowitz Department of Chemistry, University of Nebraska, Lincoln, Nebraska 12

ALTERATION OF THE REACTION PROFILE OF SERINE RACEMASE VIA SITE-DIRECTED MUTAGENESIS: IDENTIFICATION OF AN APPARENT “HOTSPOT” WITH REGARD TO REACTION MANIFOLD David L. Nelson; Gregory A. Applegate; David B. Berkowitz Department of Chemistry, University of Nebraska, Lincoln, NE

13

ENGINEERING A SPLIT-NANOLUCIFERASE REPORTER ASSAY Travis J. Nelson, Jia Zhou, and Cliff I. Stains Department of Chemistry, University of Nebraska-Lincoln 14

STRUCTURAL AND FUNCTIONAL CHARACTERIZATION OF THE NOVEL GLUTATHIONE DEGRADING ENZYME CHAC1 Michelle E. Palmer, David Beauclair, Melanie A. Simpson, Joseph J. Barycki University of Nebraska – Lincoln, Department of Biochemistry 15

BASOPHIL DEGRANULATION MARKERS TO ASSESS IGE BINDING TO BEAN GLYCOPROTEINS FOR DIAGNOSIS OF CLINICAL ALLERGENICITY Siddanakoppalu N Pramod, Afua O Tetteh, Richard E Goodman Food Allergy Research and Resource Program, Department of Food Science and Technology, University of Nebraska, Lincoln, Nebraska, USA 16

OBTAINING COMPLEMENTARY POLYPEPTIDE SEQUENCE INFORMATION FROM A SINGLE PRECURSOR ION PACKET VIA SEQUENTIAL ION MOBILITY RESOLVED ELECTRON TRANSFER AND VIBRATIONAL ACTIVATION Deepali Rathore, Forouzan Aboufazeli, and Eric D. Dodds Department of Chemistry, University of Nebraska 17

GRANULOSA CELL CYCLE REGULATION AND STEROIDOGENESIS IN A HIGH ANDROSTENEDIONE FOLLICULAR MICROENVIRONMENT Sarah M. Romereim1, Adam F. Summers2, Bill E. Pohlmeier1, Renee M. McFee1, Renata Spuri-Gomes1, Scott G. Kurz1, Anthony K. McNeel3, Robert A. Cushman3, John S. Davis4, Jennifer R. Wood1, Andrea S. Cupp1 Institutions: 1University of Nebraska–Lincoln; 2New Mexico State University, 3USDA, ARS, U.S. Meat Animal Research Center, 4University of Nebraska Medical Center 18

SERTOLI CELL-SPECIFIC ELIMINATION OF VEGFA IMPAIRS SPERMATOGONIAL STEM CELL (SSC) MAINTENANCE AND REDUCES FERTILITY IN MALE MICE Kevin M. Sargent1, Renata Spuri Gomes1, John R. Essink1, Scott G. Kurz1, William E. Pohlmeier1, Napoleone Ferrara2, Gerrit J. Bouma3, Derek J. McLean4, and Andrea S. Cupp1 1University of Nebraska-Lincoln, Lincoln, NE 2University of California-San Diego School of Medicine, San Diego, CA 3Colorado State University, Fort Collins, CO 4Phibro Animal Health Corporation, Corvallis, OR

19

COWS WITH INTRAFOLLICULAR ANDROGEN EXCESS HAVE ALTERED PATTERN OF PROGESTERONE SECRETION AND APPEAR TO BE CHRONIC OR SPORADIC ANOVULATORY Renata Spuri-Gomes, Sarah C Tenley, Mohamed A Abedal-majed, Scott G. Kurz, Jeff Bergman, Jennifer R.Wood, Andrea S. Cupp Department of Animal Science, University of Nebraska–Lincoln 20

MECHANISMS OF HEME A BIOSYNTHESIS, TRAFFICKING, AND ASSOCIATED DYSFUNCTIONS Samantha Swenson, Andrew Cannon, Oleh Khalimonchuk University of Nebraska-Lincoln, Department of Biochemistry, Redox Biology Center, Lincoln, NE 68588 21

FLUOROGENIC CHEMICAL INDUCERS OF PROTEIN OLIGOMERIZATION Bi Xu, Xinqi Zhou and Cliff I. Stains Department of Chemistry, University of Nebraska – Lincoln, Lincoln NE 68588 22

PLASMIN PROTEOLYSIS OF RECOMBINANT HUMAN FACTOR IX MADE IN TRANSGENIC PIG MILK Weijie Xu, William H. Velander Protein Purification and Characterization Lab, Department of Chemical and Biomolecular Engineering, University of Nebraska-Lincoln. 23

NANOLUCIFERASE FRAGMENTS AS SENSITIVE PROBES FOR PROTEIN SOLUBILITY IN LIVING CELLS Jia Zhao, Quyen Vu, Tiffany Truong, Travis J. Nelson & Cliff I. Stains Department of Chemistry, University of Nebraska, Lincoln, NE 24

STRUCTURES AND BIOSYNTHESIS OF PHENAZINE ANTIBIOTICS FROM LYSOBACTER ANTIBIOTICUS Yangyang Zhao1,2, Guoliang Qian2, Fengquan Liu2, Liangcheng Du1 1 Department of Chemistry, University of Nebraska-Lincoln, NE 68588; 2 College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China 25

THE 8-SILYLOXYQUINOLINE SCAFFOLD AS A VERSATILE PLATFORM FOR THE SENSITIVE DETECTION OF AQUEOUS FLUORIDE Xinqi Zhou, Rui Lai, Hui Li, and Cliff I. Stains Department of Chemistry, University of Nebraska−Lincoln, Lincoln, Nebraska 68588

Afternoon Session Disease microenvironment and Metabolic integrity

26

ANALYSIS OF FREE DRUG FRACTIONS IN SERUM BY ULTRAFAST AFFINITY EXTRACTION AND MULTI-DIMENSIONAL HIGH-PERFORMANCE AFFINITY CHROMATOGRAPHY USING IMMOBILIZED ALPHA1-ACID GLYCOPROTEIN Cong Bi, Xiwei Zheng, Sandya Beeram, and David S. Hage Department of Chemistry, University of Nebraska, Lincoln, NE 27

HYALURONAN-BASED SIGNALS MEDIATE STROMAL CELL TRANSFORMATION IN PROSTATE CANCER Christine S. Booth, Jeremy S. Payne, Caitlin O. McAtee and Melanie A. Simpson Department of Biochemistry, University of Nebraska, Lincoln, NE 28

THE ASSOCIATION OF THE GP120 V3 LOOP WITH ENVELOPE TRIMER STABILIZATION IN HIV-1 SUBTYPE C VIRUSES. Dane Bowder, Jin Wang, and Shi-hua Xiang University of Nebraska-Lincoln, Nebraska Center for Virology 29

SYNTHESIS AND CHARACTERIZATION OF ANORGANIC BONE/POLYGLYCOLIDE COMPOSITES. Lukasz Gauza, Kaitlyn Papke, Kelsey Thorpe, Jody Redepenning Department of Chemistry, University of Nebraska-Lincoln 30

ROLE OF THE RESIDENT MICROBIOTA IN PATHOBIONT-MEDIATED SUSCEPTIBILITY TO INTESTINAL INFLAMMATION J. C. Gomes Neto1, H. Kittana1, L. B. Bindels1, R. Segura Munoz1, J. M. Hostetter2 and A. E. Ramer-Tait1 1Department of Food Science and Technology, University of Nebraska-Lincoln, Lincoln, NE, USA and 2Department of Veterinary Pathology, Iowa State University, Ames, IA, USA 31

CONTRIBUTION OF ADHERENT AND INVASIVE ESCHERICHIA COLI TO GASTROINTESTINAL INFLAMMATION AND DISEASE Presenting Author : Hatem Kittana Joao Carlos Gomes Neto, Rafael Segura, Elizabeth Cody, and Amanda E. Ramer-Tait Department of Food Science and Technology, University of Nebraska-Lincoln

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MANIPULATION OF UDP-GLUCOSE DEHYDROGENASE STRUCTURE AND FUNCTION BY SMALL-MOLECULE REGULATION AND UNNATURAL AMINO ACID PHOTOCROSSLINKING George D. Grady, Jaleen Albers, Ashley Thelen, Joseph J. Barycki and Melanie A. Simpson Department of Biochemistry, University of Nebraska, Lincoln, NE 68588-0664

33

METABOLIC INVESTIGATION OF EPILEPSY AND SEIZURE ETIOLOGY FOR THE DISCOVERY OF NOVEL THERAPEUTIC AGENTS Ryan Grove1, Cory Boone1, Dana Adamcova1, Kaeli Samson2, Harrison Roundtree2, Hannah Kyllo3, Deepak Madhavan3, Kristina Simeone2, Timothy Simeone2, Jiri Adamec1 1Department of Biochemistry, University of Nebraska Lincoln; 2Creighton University, Omaha, NE; 3University of Nebraska Medical Center, Omaha, NE.

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CANDIDA ALBICANS QUORUM SENSING MOLECULE STIMULATES THE MIGRATION OF INNATE IMMUNE CELLS IN VIVO. Jessica C. Hargarten1, Tyler C. Moore1,2, Deborah M. Brown1,3, Thomas M. Petro2, Kenneth W. Nickerson1, and Audrey L. Atkin.1 School of Biological Sciences1, Department of Oral Biology2, and Nebraska Center for Virology3, University of Nebraska, Lincoln, Nebraska.

35

TARGETED DELIVERY OF MICRORNA BY ENGINEERED LIPID NANOPARTICLES FOR THE TREATMENT OF BREAST CANCER Stephen L. Hayward and Srivatsan Kidambi Department of Chemical and Biomolecular Engineering, University of Nebraska-Lincoln, NE

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STABILIZATION OF OUTER DOMAIN OF GP120 FOR HIV-1 VACCINE IMMUNOGEN DESIGN Duoyi Hu*, Jesse Thompson*, Pankaj Kumar*, Jizu Yi, Dane Bowder, Charles Wood, Mark Wilson and Shi-hua Xiang Nebraska Center for Virology, University of Nebraska-Lincoln

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A BACTERIAL EFFECTOR THAT USES CALMODULIN TO TARGET THE PLANT MICROTUBULE NETWORK Panya Kim1,3, Ming Guo1,2, Guangyong Li1,2, Christian G. Elowsky4, and James R. Alfano 1,2 1 Center for Plant Science Innovation, University of Nebraska, Lincoln, NE 68588-0660 2 Department of Plant Pathology, University of Nebraska, Lincoln, NE 68588-0722 3 School of Biological Sciences, University of Nebraska, Lincoln, NE 68588-0118 4 Center for Biotechnology, University of Nebraska, Lincoln, NE 68588-0665

38

ANALYSIS OF DRUG-PROTEIN INTERACTIONS DURING DIABETES BY HIGH PERFORMANCE AFFINITY CHROMATOGRAPHY

Zhao Li, Ryan Matsuda, David S. Hage Department of chemistry, University of Nebraska-Lincoln 39

FREE FRACTION ANALYSIS BY DISPLACEMENT ASSAY BASED ON HIGH PERFORMANCE AFFINITY CHROMATOGRAPHY Elli Kaufman, Ryan Matsuda, Xiwei Zheng and David. S. Hage University of Nebraska-Lincoln Chemistry Department and UNL UCARE Program 40

PROSTATE TUMOR CELL EXOSOMES CONTAINING HYALURONIDASE HYAL1 STIMULATE THE MOTILITY OF PROSTATE STROMAL CELLS Caitlin O. McAtee1, Terri Fangman2, Oleh Khalimonchuk1, Steve Caplan3,4, Melanie A. Simpson1,4

1 Department of Biochemistry, University of Nebraska, Lincoln, NE 2 Morrison Microscopy Facility, University of Nebraska, Lincoln, NE 3 Department of Biochemistry and Molecular Biology, University of Nebraska Medical Center, Omaha, NE 4 Fred and Pamela Buffett Cancer Center, Omaha, NE 41

CLEARANCE SYSTEMIC HEPARIN VIA STAB-2/HARE IN THE LIVER SINUSOIDAL ENDOTHELIAL Colton Miller and Edward Harris Department of Biochemistry, University of Nebraska-Lincoln 42

COMPUTATIONAL SYSTEMS BIOLOGY EFFORTS TO BETTER UNDERSTAND DISEASES Akram Mohammed1, Nicholas Galt1, Bryan Kowal1, David Tichy1, Greyson Biegert1, Travis Schreier1, Yizi Mao2, Tomas Helikar1 1Department of Biochemistry, 2School of Biological Sciences, University of Nebraska 43

SUBSTRATE ELASTICITY REGULATES PRIMARY HEPATOCYTES FUNCTIONS Vaishaali Natarajan 1, Eric J Berglund 1, Dorothy X Chen 1, Srivatsan Kidambi 1,2,3 1 Department of Chemical and Biomolecular Engineering, University of Nebraska-Lincoln, NE 2 Nebraska Center for Materials and Nanoscience, University of Nebraska-Lincoln, NE, 3 Regenerative Medicine Program, University of Nebraska Medical Center, NE 44

INDIVIDUAL AND COMBINED ADMINISTRATION OF GALACTOOLIGOSACCHARIDE AND BIFIDOBACTERIA ADOLESCENTIS IVS-1 TO HIGH FAT-FED MICE CAUSES DIFFERENTIAL ADAPTATION OF THE MICROBIOME AND METABOLIC PROFILE. Rafael Segura, Laure Bindels, Janina Krumbeck, Carlos Gomes, Hatem Kittana, Maria Quintero, Elizabeth Cody, Jens Walter and Amanda E. Ramer-Tait. Department of Food Science and technology, University of Nebraska, Lincoln, NE

45

QUANTITATIVE HIGH PERFORMANCE AFFINITY CHROMATOGRAPHY ANALYSIS OF INTERACTIONS OF 8-ANILINO-1-NAPHTHALENESULFONIC ACID WITH HSA AND GLYCATED HSA D. Suresh1,2, Zhao Li1, David S Hage1 1University of Nebraska – Lincoln, NE 68588, USA, 2Tumkur University, Tumkur, Karnataka 572103, India 46

INTERFERON REGULATORY FACTOR 3 DEFICIENCY CONTRIBUTES TO IMPAIRED MEMORY T CELL FUNCTION IN RESPONSE TO INFLUENZA INFECTION Alexander J. Vogel1,2 Tyler C. Moore1,2 Thomas M. Petro3 and Deborah M. Brown1,2 1 School of Biological Sciences 2 Nebraska Center for Virology, University of Nebraska-Lincoln 3 Department of Oral Biology, University of Nebraska Medical Center 47

INCREASED FIRMICUTES IN THE CECUM OF OBESE FEMALE MICE IS CORRELATED WITH INCREASED POU5F1 AND DPPA3 MRNAS IN GROWING OOCYTES Fang Xie1, Christopher L. Anderson2, Kelsey Timme2, Samodha C. Fernando2, and Jennifer R. Wood1 Department of Animal Science, University of Nebraska, Lincoln, NE 48

REGULATORS OF SPHINGOLIPID BIOSYNTHESIS MODULATE PLANT IMMUNITY Fan Yang, Athen Kimberlin, Edgar Cahoon, and Jim Alfano Center for Plant Science Innovation, University of Nebraska 49

INVESTIGATING THE ROLE OF UDP-GLUCOSE DEHYDROGENASE IN MAINTAINING ANDROGEN SENSITIVITY IN PROSTATE TUMOR DEVELOPMENT Brenna M. Zimmer, Eileen Loughman, and Melanie A. Simpson Department of Biochemistry, University of Nebraska, Lincoln, NE 50

UDP-GLUCURONATE PARTITIONING IN METABOLIC DETOXIFICATION Michelle E. Palmer, Jonathan E. Markham, Joseph J. Barycki, and Melanie A. Simpson Department of Biochemistry, University of Nebraska, Lincoln, NE    

NOTES