Generating Enzyme And Radical‐Mediated Bisubstrates As Tools For Investigating Gcn5‐Related N‐Acetyltransferases,
2017
Loyola University Chicago
Generating Enzyme And Radical‐Mediated Bisubstrates As Tools For Investigating Gcn5‐Related N‐Acetyltransferases, Cory T. Reidl, Karolina A. Majorek, Joseph Dang, David Tran, Kristen Jew, Melissa Law, Yasmine Payne, Wladek Minor, Daniel P. Becker, Misty L. Kuhn
Chemistry: Faculty Publications and Other Works
Gcn5‐related N‐acetyltransferases (GNATs) are found in all kingdoms of life and catalyze important acyl transfer reactions in diverse cellular processes. While many 3D structures of GNATs have been determined, most do not contain acceptor substrates in their active sites. To expand upon existing crystallographic strategies for improving acceptor‐bound GNAT structures, we synthesized peptide substrate analogs and reacted them with CoA in PA4794 protein crystals. We found two separate mechanisms for bisubstrate formation: (a) a novel X‐ray induced radical‐mediated alkylation of CoA with an alkene peptide and (b) direct alkylation of CoA with a halogenated peptide. Our approach is widely …
An Approach To Identify Mycobacteriophage Diversity Prior To Dna Sequencing,
2017
Western Kentucky University
An Approach To Identify Mycobacteriophage Diversity Prior To Dna Sequencing, Charles Gregory
Mahurin Honors College Capstone Experience/Thesis Projects
Over 6,869 Mycobacteriophages have been isolated and purified. Of these, 1,367 genomes have been sequenced at the DNA level and more are added each year through the SEA-PHAGES program. Sequenced mycobacteriophages are grouped into clusters based on a 50% or greater nucleotide identity. The number and breadth of these clusters represents the diversity present in the environment. Each year, as new phages are discovered by students in the SEA-PHAGES program, the question arises, “Which isolates should we sequence?” In order to sequence phages that represent the greatest possible diversity, and thus broaden under-represented clusters and identify new singletons, we need …
Investigating The Synergistic Effects Of Cisplatin And Two Curcuminoid Compounds On Cancer,
2017
Western Kentucky University
Investigating The Synergistic Effects Of Cisplatin And Two Curcuminoid Compounds On Cancer, Denis Hodzic
Mahurin Honors College Capstone Experience/Thesis Projects
Cisplatin is an anti-cancer drug effective against several cancers which can produce the serious side-effect of hearing loss. Curcumin, a natural plant compound, can increase the activity of cisplatin against cancer and counteract cisplatin’s effect against hearing. Because curcumin exhibits poor bioavailability, there is considerable interest in developing synthetic curcumin analogs (curcuminoids) that are more soluble and which retain anti-cancer activity and otoprotective function. This study investigated whether two curcuminoids, EF-24 and CLEFMA, increase the cytotoxic and ototoxic effects of cisplatin against the lung cancer cell line, A549, and the colorectal cancer cell line, Caco2. Cytotoxicity was measured by using …
The General Amino Acid Permease Gap1 Is Regulated Differentially By Torc1 Activation And Inhibition,
2017
Duquesne University
The General Amino Acid Permease Gap1 Is Regulated Differentially By Torc1 Activation And Inhibition, Ray Bowman
D.U.Quark
How does cell signaling in response to extracellular stressors impact the trafficking of membrane proteins? In particular, the TORC1 complex plays a key role in this process and while some details of this system have reported, in a recent Journal of Biological Chemistry publication, Andre’s group has revealed new details of this pathway focusing on the general amino acid permease Gap1 as a model cargo. Andre et al. describe a novel and distinct pathway wherein ubiquitylation and downregulation of Gap1 is regulated not only by amino acid-induced activation of TORC1, but also by numerous sources of TORC1 inhibition and cellular …
Spatiotemporal Regulation Of Atg1 Kinase Activation In Selective Autophagy,
2017
Duquesne University
Spatiotemporal Regulation Of Atg1 Kinase Activation In Selective Autophagy, Ning Sun
D.U.Quark
Autophagy is a potent intracellular degradation system and thus its activation requires exquisite regulation to maintain cellular homeostasis. Atg1, a serine-threonine protein kinase, is essential in both selective and non-selective autophagy. New findings suggest that in selective autophagy, Atg1 is activated at the vacuole by convergence of two independent recruitment pathways to prevent aberrant autophagy induction.
Hsp70 Conformational Plasticity Allows For Expansive Chaperone Role,
2017
Duquesne University
Hsp70 Conformational Plasticity Allows For Expansive Chaperone Role, Megan Bean
D.U.Quark
The Hsp70 system is an essential component of chaperone activity in many organisms. Hsp70 functions include: protein folding, aggregation prevention, trafficking, and enzyme regulation. Hsp70’s ability to bind such a vast array of substrates suggests wide range of conformational plasticity. By utilizing a single mode optical tweezers technique, Mashaghi1 et al., confirms previous theories Hsp70 binds and stabilizes extended peptide segments but also partially folded and near-native protein.
Plasmodium Falciparum 26s Proteasome Network: A Mystery Solved,
2017
Duquesne University
Plasmodium Falciparum 26s Proteasome Network: A Mystery Solved, Christina Grogan
D.U.Quark
One ofthe most devastating diseases thatthreatens the world population is malaria. The 26S proteasome complex of the malaria parasite Plasmodium falciparum, which was previously unknown, was characterized by the authors through an affinity purification protocol that isolated functional 26S proteasome complexes. This allowed for the identification of subunit composition and PfUSP14, a proteasomeassociated deubiquitinase. This new understanding presents a potential target to disrupt protein regulation in thequest for effective antimalarial strategies.
Autophagy Inhibition In Pain: Role Of A Microrna,
2017
Duquesne University
Autophagy Inhibition In Pain: Role Of A Microrna, Andrea Stevens
D.U.Quark
Neuropathic pain caused by peripheral nerve injury (PNI) leads to the activation and infiltration of microglial cells and to a neuroinflammatory-induced pain state. miRNAs and autophagy are two main factors and/or mechanisms which have the ability to alter the pain state. In this study, miR-195 was shown to be markedly increased after PNI and associated with the pain phenotype. In addition, inhibition of autophagy in vivo led to p62 accumulation, decreased production of LC3, and inhibition of ATG14.
Inhibiting Translesion Dna Synthesis As An Approach To Combat Drug Resistance To Dna Damaging Agents,
2017
Cleveland State University
Inhibiting Translesion Dna Synthesis As An Approach To Combat Drug Resistance To Dna Damaging Agents, Jung-Suk Choi, Seol Kim, Edward Motea, Anthony J. Berdis
Chemistry Faculty Publications
Anti-cancer agents exert therapeutic effects by damaging DNA. Unfortunately, DNA polymerases can effectively replicate the formed DNA lesions to cause drug resistance and create more aggressive cancers. To understand this process at the cellular level, we developed an artificial nucleoside that visualizes the replication of damaged DNA to identify cells that acquire drug resistance through this mechanism. Visualization is achieved using "click" chemistry to covalently attach azide-containing fluorophores to the ethynyl group present on the nucleoside analog after its incorporation opposite damaged DNA. Flow cytometry and microscopy techniques demonstrate that the extent of nucleotide incorporation into genomic DNA is enhanced …
Interleukin-18 Polymorphism As An Inflammatory Index In Metabolic Syndrome: A Preliminary Study,
2017
Aga Khan University
Interleukin-18 Polymorphism As An Inflammatory Index In Metabolic Syndrome: A Preliminary Study, Syeda Sadia Fatima, Zehra Jamil, Syed Hani Abidi, Daniyal Nadeem, Zara Bashir, Ahmed Ansari
Department of Biological & Biomedical Sciences
Aim: To assess circulatory levels of interleukin-18 (IL-18) and determine whether the presence of IL-18 promoter polymorphism influences metabolic syndrome phenotypes.
Methods: This study recruited one hundred and eighty individuals divided into three groups with sixty subjects each as: Normal weight (18.0-22.9 kg/m2), overweight (23.0-25.9 kg/m2) and obese (> 26.0 kg/m2) according to South Asian criteria of BMI. Fasting blood glucose (FBG), Lipid profile, insulin, IL-18 and tumor necrosis factor (TNF)alpha were measured using ELISA kits, whereas low density lipoprotein (LDL)-cholesterol, insulin resistance (HOMA-IR) and insulin sensitivity (QUICKI) were calculated. The body fat percentage (BF) was measured through bioelectrical impedance …
Exploring Cancer Metabolism Using Stable Isotope-Resolved Metabolomics (Sirm),
2017
University of Kentucky
Exploring Cancer Metabolism Using Stable Isotope-Resolved Metabolomics (Sirm), Ronald C. Bruntz, Andrew N. Lane, Richard M. Higashi, Teresa W. -M. Fan
Center for Environmental and Systems Biochemistry Faculty Publications
Metabolic reprogramming is a hallmark of cancer. The changes in metabolism are adaptive to permit proliferation, survival, and eventually metastasis in a harsh environment. Stable isotope-resolved metabolomics (SIRM) is an approach that uses advanced approaches of NMR and mass spectrometry to analyze the fate of individual atoms from stable isotope-enriched precursors to products to deduce metabolic pathways and networks. The approach can be applied to a wide range of biological systems, including human subjects. This review focuses on the applications of SIRM to cancer metabolism and its use in understanding drug actions.
Growth Trade-Offs Accompany The Emergence Of Glycolytic Metabolism In Shewanella Oneidensis Mr-1,
2017
Harvard University
Growth Trade-Offs Accompany The Emergence Of Glycolytic Metabolism In Shewanella Oneidensis Mr-1, Lon Chubiz, Christopher Marx
Biology Department Faculty Works
Bacteria increase their metabolic capacity via the acquisition of genetic material or by the mutation of genes already present in the genome. Here, we explore the mechanisms and trade-offs involved when Shewanella oneidensis, a bacterium that typically consumes small organic and amino acids, rapidly evolves to expand its metabolic capacity to catabolize glucose after a short period of adaptation to a glucose-rich environment. Using whole-genome sequencing and genetic approaches, we discovered that deletions in a region including the transcriptional repressor (nagR) that regulates the expression of genes associated with catabolism of N-acetylglucosamine are the common basis for evolved glucose metabolism …
Current Status Of Helicobacter Pylori Association With Haematological And Cardiovascular Diseases: A Mini Review,
2017
Aga Khan University
Current Status Of Helicobacter Pylori Association With Haematological And Cardiovascular Diseases: A Mini Review, Jibran Sualeh Muhammad, Syed Faisal Zaidi, Sheikh Abdul Saeed, Muhammad Ishaq
Department of Biological & Biomedical Sciences
Helicobacter pylori infection is considered the most commonly prevalent gastrointestinal pathogen where it manages to survive despite the hostile environment of human stomach, leading to various gastric diseases including gastric cancer. Due to the chronic inflammatory state induced by H. pylori and its interaction with host immune system have diverted researchers to investigate its correlation with systemic diseases outside of the gastrointestinal tract. This literature review was done to explore the association of H. pylori infection with haematological and cardiovascular diseases. We used medical subject heading (MeSH) terms "Helicobacter pylori" with "inflammation," "haematological diseases," "coronary heart diseases" or "vascular diseases" …
Anthrax, Matrix Biology, And Angiogenesis: Capillary Morphogenesis Gene 2 Mediates Activity And Uptake Of Type Iv Collagen-Derived Anti-Angiogenic Peptides,
2017
Brigham Young University
Anthrax, Matrix Biology, And Angiogenesis: Capillary Morphogenesis Gene 2 Mediates Activity And Uptake Of Type Iv Collagen-Derived Anti-Angiogenic Peptides, Jordan Grant Finnell
Theses and Dissertations
Capillary Morphogenesis Gene 2 (CMG2) is a type I transmembrane, integrin-like receptor. It was originally identified as one of several genes upregulated during capillary formation. It was subsequently identified as one of two physiological anthrax toxin receptors, where CMG2 serves as a cell-surface receptor for anthrax toxin and mediates entry of the toxin into cells via clathrin-dependent endocytosis. Additionally, loss-of-function mutations in CMG2 cause the genetic disorder hyaline fibromatosis syndrome (HFS), where the core symptom is dysregulation of extracellular matrix homeostasis (ECM), including excessive accumulation of proteinaceous hyaline material; HFS clearly indicates that CMG2 plays an essential function in ECM …
Enzyme Encapsulation, Biosensing Endocrine Disrupting Chemicals, And Bio-Therapeutic Expression Platforms Using Cell-Free Protein Synthesis,
2017
Brigham Young University
Enzyme Encapsulation, Biosensing Endocrine Disrupting Chemicals, And Bio-Therapeutic Expression Platforms Using Cell-Free Protein Synthesis, Seung Ook Yang
Theses and Dissertations
Cell-free protein synthesis (CFPS) is a powerful protein expression platform where protein synthesis machinery is borrowed from living organisms. Target proteins are synthesized in a reaction tube together with cell extract, amino acids, energy source, and DNA. This reaction is versatile, and dynamic optimizations of the reaction conditions can be performed. The "œopen" nature of CFPS makes it a compelling candidate for many technologies and applications. This dissertation reports new and innovative applications of CFPS including 1) enzyme encapsulation in a virus-like particle, 2) detection of endocrine disrupting chemicals in the presence of blood and urine, and 3) expression of …
The Mechanistic Requirements Of Passive H+ Import Through The Na, K-Atpase,
2017
Illinois State University
The Mechanistic Requirements Of Passive H+ Import Through The Na, K-Atpase, Kevin S. Stanley
Theses and Dissertations
This work focuses on the elucidation of the mechanism of passive proton import through the Na,K-ATPase. This enzyme uses the energy in ATP hydrolysis to exchange three intracellular Na+ for two extracellular K+ to maintain ion gradients within the cell, and while in the absence of physiological external Na+ and K+, the phosphorylated externally open (E2P) conformation passively imports protons, generating an inward current (IH). Chapter one reports on the effects of intracellular cations and nucleotides to shift the Na,K-ATPase into the E2P conformation. We identified that a combination of either internal Na+ and ATP or K+ and Pi. In …
A Novel Population Of Cardiovascular Progenitors Persist In Neonates As Mesendodermal Cells,
2017
Loma Linda University
A Novel Population Of Cardiovascular Progenitors Persist In Neonates As Mesendodermal Cells, Julia Kim
Loma Linda University Electronic Theses, Dissertations & Projects
The rise in mortality due to cardiovascular disease has increased the need to develop an efficient regenerative therapeutic for heart failure. Numerous cell-based therapies have been investigated for myocardial regeneration; however, an optimal progenitor has yet to be discovered. Identifying a resident cell population with enhanced ability to differentiate into multiple lineages would greatly contribute to the field of stem cell-based regenerative therapy. Evidence suggests that endogenous cardiovascular progenitor cells (CPCs) that have been isolated from the heart itself express ISL1, KDR, and MESP1, and are capable of differentiating into all major cardiac lineages. The earlier developmental stage at which …
2d Nmr Structural Study Of A Dna Duplex Containing Two Unnatural Base Pairs,
2017
University of San Diego
2d Nmr Structural Study Of A Dna Duplex Containing Two Unnatural Base Pairs, Weston Preising
Undergraduate Honors Theses
DNA is the genetic blueprint responsible for the traits of living organisms. The "genetic alphabet" consists of four molecules called nucleobases, represented by the letters A, T, G, and C strung together in a "strand." A DNA duplex consists of two "strands" held together such that A is across from T and G across from C, referred to A-T and G-C "base pairs". Recently, the genetic alphabet or code has been expanded by synthetic biologists in an effort to unveil new insights to the machinery of DNA replication, and potentially develop new medicines to cure diseases like HIV. The new …
Rapid Reversible Borane To Boryl Hydride Exchange By Metal Shuttling On The Carborane Cluster Surface,
2017
University of South Carolina
Rapid Reversible Borane To Boryl Hydride Exchange By Metal Shuttling On The Carborane Cluster Surface, Bennett J. Eleazer, Mark D. Smith, Alexey A. Popov, Dmitry V. Peryshkov
Faculty Publications
In this work, we introduce a novel concept of a borane group vicinal to a metal boryl bond acting as a supporting hemilabile ligand in exohedrally metalated three-dimensional carborane clusters. The (POBOP)Ru(Cl)(PPh3) pincer complex (POBOP ¼ 1,7-OP(i-Pr)2-m-2-carboranyl) features extreme distortion of the two-center-two-electron Ru–B bond due to the presence of a strong three-centertwo-electron B–H/Ru vicinal interaction. Replacement of the chloride ligand with a hydride afforded the (POBOP)Ru(H)(PPh3) pincer complex, which possesses B–Ru, B–H/Ru, and Ru–H bonds. This complex was found to exhibit a rapid exchange between hydrogen atoms of the borane and the terminal hydride through metal center shuttling between …
Estrogen-Activated Mdm2 Disrupts Mammary Tissue Architecture Through A P53-Independent Pathway,
2017
CUNY Hunter College
Estrogen-Activated Mdm2 Disrupts Mammary Tissue Architecture Through A P53-Independent Pathway, Nandini Kundu, Angelika Brekman, Jun Yeob Kim, Gu Xiano, Chong Gao, Jill Bargonetti
Publications and Research
The Cancer Genome Atlas (TCGA) data indicate that high MDM2 expression correlates with all subtypes of breast cancer. Overexpression of MDM2 drives breast oncogenesis in the presence of wild-type or mutant p53 (mtp53). Importantly, estrogen-receptor positive (ER+) breast cancers overexpress MDM2 and estrogen mediates this expression. We previously demonstrated that this estrogen-MDM2 axis activates the proliferation of breast cancer cell lines T47D (mtp53 L194F) and MCF7 (wild-type p53) in a manner independent of increased degradation of wildtype p53 (ie, p53-independently). Herein we present data supporting the role of the estrogen-MDM2 axis in regulating cell proliferation and mammary tissue architecture of …
