Chemical Biology Progress & Challenges May 4th, 2013 College Park, Maryland Sixth Annual Frontiers at the Chemistry‐Biology Interface Symposium University of Maryland College Park, May 4th, 2013 Organizers Herman O. Sintim and Shuwei Li (University of Maryland, College Park) Jim Fishbein (University of Maryland, Baltimore County) John Koh and Millie Sullivan (University of Delaware) Steve Rokita and Jin Zhang (Johns Hopkins University) E. James Petersson (University of Pennsylvania) Ronen Marmorstein (The Wistar Institute) Sponsors Department of Chemistry The Wistar Institute University of Pennsylvania Department of Chemistry and Biochemistry University of Delaware School of Medicine, Department of Pharmacology Johns Hopkins University Department of Chemistry Chemistry-Biology Interface Training Program John Hopkins University Johns Hopkins University Department of Chemistry and Biochemistry University of Maryland, Baltimore County Department of Chemistry and Biochemistry University of Maryland, College Park ACS Chemical Biology 1 Sixth Annual Frontiers at the Chemistry‐Biology Interface Symposium University of Maryland College Park, May 4th, 2013 Program Schedule 8:00-9:00 Pick up name tag, breakfast and set up posters 9:00-10:45 AM, Session I 9:00-9:05 am Introduction byChair: UMD DepartmentJames Petersson Chair, (University Michael Doyle of Pennsylvania) 9:00-9:05 Welcome Message by UMD Department Chair, Michael Doyle 9:05-9:10 Introduction by Chair of the Organizing Committee, Herman Sintim 9:10-9:35 Ben L. Feringa (University of Groningen) Exploring Chiral Space in Asymmetric Catalysis 9:35-10:00 Alex Deiters (North Carolina State University) Synthetic Chemical Tools for the Regulation of Cellular Processes 10:00-10:20 Jason Kahn (Poster speaker, University of Maryland College Park) Rationally Designed Coiled‐coil DNA Looping Peptides Control DNA Topology 10:20-10:45 Barbara Gerratana (National Institute of Health) Navigating the NIH Funding Process 10:45-11:00 AM, Coffee Break 11:00-12:15 PM, Session II Chair: Catherine Grimes (University of Delaware) 11:00-11:25 David Chenoweth (University of Pennsylvania) Structure Specific Nucleic Acid Targeting 11:25-11:50 Bruce Yu (University of Maryland Baltimore) Develop Fluorinated Dendrimers for 19F MRI 11:50-12:15 Tanakari Inoue (John Hopkins University) Total Synthesis of Chemotaxis 12:15-1:50 PM, Lunch and Poster 1:50-3:25 PM, Session III Chair: Herman Sintim (University of Maryland, College Park) 1:50-2:15 Cynthia Dowd (George Washington University) Dxr Inhibitors to Combat Mycobacterium tuberculosis 2:15-2:40 Catherine Grimes (University of Delaware) Chemical Tools to Study the Activation of the Intracellular Innate Immune Protein Nod2 2:40-3:00 Meiyao Wang (Poster speaker, National Institute of Standards and Technology) Absolute Quantification and Isoforms Differentiation of Membrane‐Associated Proteins Using Multiple Reaction Monitoring Mass Spectrometry and Isotope Labeled Full‐Length Protein Standards 3:00-3:25 Christian Melander (North Carolina State University) Small Molecule Suppression of Antibiotic Resistance 2 Sixth Annual Frontiers at the Chemistry‐Biology Interface Symposium University of Maryland College Park, May 4th, 2013 3:25-3:40 PM, Coffee Break 3:40-5:30 PM, Session IV Chair: Shuwei Li (University of Maryland, College Park) 3:40-4:05 Hui Zhang (Johns Hopkins University) Glycomic Analysis Using Solid-Phase Glycan Extraction and Mass Spectrometry 4:05-4:30 Jeff Gildersleeve (National Cancer Institute) Carbohydrate-Binding Serum Antibodies as Biomarkers for Personalized Medicine 4:30-5:30 Gerald Hart (Keynote Speaker, John Hopkins University) Glycomics Reveals Extensive Crosstalk Between O-GlcNAcylation and Other PTMs: Roles in Signaling, Transcription and Chronic Disease 5:30-5:40 PM, Presentation of Poster Prizes and Closing Remarks Herman Sintim (University of Maryland, College Park) 3 Sixth Annual Frontiers at the Chemistry‐Biology Interface Symposium University of Maryland College Park, May 4th, 2013 Posters (in alphabetic order by the presenter's last name) 1. Presenter: Ara Abramyan, University of the Sciences Complete List of Authors: Ara Abramyan, Zhiwei Liu, Vojislava Pophristic Title: "Molecular Dynamics Investigation of DNA-binding Foldamers" "Foldamers are synthetically derived oligomers inspired by the structures of biopolymers. Some of these oligomers have been programmed to selectively recognize specific sequences in DNA. A number of human cancers are caused by dysregulation of transcription factors leading to an anomalous gene expression. Therefore ligands that are capable of binding DNA sequences and interrupting transcription factor-DNA interactions are of a great interest. We apply molecular dynamics methods to study oligoamides experimentally shown to bind to the DNA minor groove. We analyze structural changes in DNA upon oligoamide binding as well as the influence of various chemical and structural changes in a series of oligoamides. Particularly, we discuss how the presence and position of positively charged amino group on oligoamides' turn unit perturbs the DNA sequence of interest. Our aim is to design an oligoamide with improved affinity and selectivity for further experimental studies. " 2. Presenter: Smriti Agrawal, University of Delaware Complete List of Authors: "Smriti A. Agrawal, Atul Kakrana, David Scheiblin, Christine A. Dang, Stephanie M. Waters, Abhyudai Singh, Hozumi Motohashi, Salil A. Lachke" Title: "Deficiency of small Maf family transcription factors MafG and MafK disrupts gene regulation in the lens and causes cataract" "The ocular lens is a transparent tissue that functions to focus light on the retina and is critical for providing us with high-resolution vision. Loss of lens transparency, which occurs due to genetic changes or aging, results in a disease called “cataract” – the leading cause of blindness worldwide. To devise strategies to delay or prevent cataract formation, it is critical to first understand how the lens develops and maintains transparency. We used a bioinformatics approach, iSyTE (integrated System Tool for Eye gene discovery) to identify the small Maf (musculoaponeurotic fibrosarcoma) family gene MafG as a potential regulator in the lens. The Maf gene family encodes basic leucine zipper transcription regulators that are classified into “large” and “small” MAF subgroups. Although mutations in the large Maf family gene Maf (c-Maf) are associated with human congenital cataracts, the function of the small Maf subgroup proteins MafG, MafK, and MafF in the lens remains uncharacterized. Our expression analysis confirmed iSyTE’s prediction that MafG is highly enriched in both embryonic and adult lens tissue. We also detected MafK expression in the lens although at significantly lower levels. To test the function of these small Maf genes in the lens, we generated and characterized mouse mutants that carry various combinations of MafG and MafK null alleles. These analyses demonstrate that Mafg-/-:Mafk+/- compound mutants exhibit severe lens defects starting at 2 months and develop distinct pre-senile cataract at age 6 months. To identify the target genes of these transcriptional regulators, we performed whole genome transcript profiling by microarrays on lenses from control and Mafg-/-:Mafk+/- mutant mice. These analyses indicate that expression of the heat shock protein gene Hspb1(Hsp27), a heme oxygenase gene Hmox1(Hsp32) and a novel gene Ttc27 are differentially regulated in Mafg-/-:Mafk+/- compound mutant lenses. Interestingly, Hspb1 is found to directly interact with and stabilize lens crystallin proteins Crystallin alpha A and Crystallin alpha B, which are necessary for lens transparency. Hspb1 is also found to be downregulated in mouse lenses carrying homozygous null mutations of the RNA granule component gene Tdrd7, deficiency of which in mouse or human causes cataract. Collectively, our data have identified and characterized a new role for MafG and MafK in controlling lens fiber cell gene expression, and demonstrated that deficiency of these proteins in mouse results in cataract. " 3. Presenter: Solongo Batjargal, University of Pennsylvania Complete List of Authors: "Solongo Batjargal, E. James Petersson" Title: "Labeling Proteins with Fluorophore/Thioamide FRET Pairs" 4 Sixth Annual Frontiers at the Chemistry‐Biology Interface Symposium University of Maryland College Park, May 4th, 2013 "A thioamide, the substitution of a carbonyl oxygen with a sulfur in the peptide backbone, is capable of quenching the fluorescence of p-cyanophenylalanine (Cnf) in a distant dependent fashion. The combination of native chemical ligation and unnatural amino acid (UAA) mutagenesis can be used to generate a thioamide/Cnf FRET pair in a protein. We recently have shown that a thioamide containing full-sized protein can be generated by ligating N-terminal Cys protein expressed in E. Coli and thioamide-containing peptide thioester. This method locates the thioamide in the N-terminal region of the protein. To insert thioamides at the C-terminus, we demonstrate that the intein fusion constructs can be employed to generate a protein thioester. The use of UAA mutagenesis and intein fusion permits us to obtain a Cnf-labeled protein thioester fragment which then can be ligated to a thioamide- labeled peptide synthesized on solid phase. This combination of methods allows for quick access to double-labeled proteins with site-specificity." 4. Presenter: Petrina Boucher, University of Maryland Complete List of Authors: