Comparative Analysis of Mutants in the Mycothiol Biosynthesis Pathway in Mycobacterium Smegmatis
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A Putative Cystathionine Beta-Synthase Homolog of Mycolicibacterium Smegmatis Is Involved in De Novo Cysteine Biosynthesis
University of Arkansas, Fayetteville ScholarWorks@UARK Theses and Dissertations 5-2020 A Putative Cystathionine Beta-Synthase Homolog of Mycolicibacterium smegmatis is Involved in de novo Cysteine Biosynthesis Saroj Kumar Mahato University of Arkansas, Fayetteville Follow this and additional works at: https://scholarworks.uark.edu/etd Part of the Cell Biology Commons, Molecular Biology Commons, and the Pathogenic Microbiology Commons Citation Mahato, S. K. (2020). A Putative Cystathionine Beta-Synthase Homolog of Mycolicibacterium smegmatis is Involved in de novo Cysteine Biosynthesis. Theses and Dissertations Retrieved from https://scholarworks.uark.edu/etd/3639 This Thesis is brought to you for free and open access by ScholarWorks@UARK. It has been accepted for inclusion in Theses and Dissertations by an authorized administrator of ScholarWorks@UARK. For more information, please contact [email protected]. A Putative Cystathionine Beta-Synthase Homolog of Mycolicibacterium smegmatis is Involved in de novo Cysteine Biosynthesis A thesis submitted in partial fulfillment of the requirement for the degree of Master of Science in Cell and Molecular Biology by Saroj Kumar Mahato Purbanchal University Bachelor of Science in Biotechnology, 2016 May 2020 University of Arkansas This thesis is approved for recommendation to the Graduate Council. ___________________________________ Young Min Kwon, Ph.D. Thesis Director ___________________________________ ___________________________________ Suresh Thallapuranam, Ph.D. Inés Pinto, Ph.D. Committee Member Committee Member ABSTRACT Mycobacteria include serious pathogens of humans and animals. Mycolicibacterium smegmatis is a non-pathogenic model that is widely used to study core mycobacterial metabolism. This thesis explores mycobacterial pathways of cysteine biosynthesis by generating and study of genetic mutants of M. smegmatis. Published in vitro biochemical studies had revealed three independent routes to cysteine synthesis in mycobacteria involving separate homologs of cysteine synthase, namely CysK1, CysK2, and CysM. -
Great South African Molecules: the Case for Mycothiol
REVIEW ARTICLE C.M. Nkambule, 67 S. Afr. J. Chem., 2017, 70, 67–81, <http://journals.sabinet.co.za/sajchem/>. Great South African Molecules: The Case For Mycothiol Comfort M. Nkambule Department of Chemistry, Tshwane University of Technology, Pretoria, 0001, South Africa. Received 13 September 2016, revised 15 March 2017, accepted 17 March 2017. ABSTRACT South Africa has one of the oldest chemical societies in the world and has a long history of natural products, synthetic, and medici- nal chemistry yet the visibility of molecules discovered or synthesized in South Africa is very low. Is this because South African scientists are incapable of discovering influential and celebrated molecules, or is there inadequate publicity of such discoveries? Perspective profiles on the discovery and scope of research on ‘Great South African Molecules’ should be a good start to redress this state of affairs. One such molecule deserving publicity is the antioxidant mycothiol which is produced by mycobacteria. This is a molecule of interest not only because of its medicinal potential in the fight against tuberculosis, but also from synthetic methodology and enzyme inhibition studies. This review aims to illuminate the scope of research in mycothiol chemistry for the purpose of promoting multidisciplinary investigations related to this South African molecule. KEYWORDS Mycothiol, Mycobacterium tuberculosis, great molecules, chemistry Nobel Prize. Table of Contents 1. What is a Great Molecule?· · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · 67 2. Is Mycothiol a Great Molecule? · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · 71 2.1. Is Mycothiol a South African Molecule? · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · 71 3. Why is Mycothiol a Molecule of Great Potential? · · · · · · · · · · · · · · · · · · · · · · · · · · · 72 3.1. Mycothiol’s Significance to the TB Problem· · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · 72 3.2. -
Biosynthesis and Functions of Bacillithiol, a Major Low-Molecular-Weight Thiol in Bacilli
Biosynthesis and functions of bacillithiol, a major low-molecular-weight thiol in Bacilli Ahmed Gaballaa,1, Gerald L. Newtonb,1, Haike Antelmannc, Derek Parsonaged, Heather Uptone, Mamta Rawate, Al Claiborned, Robert C. Faheyb, and John D. Helmanna,2 aDepartment of Microbiology, Cornell University, Ithaca, NY 14853-8101; bDepartment of Chemistry and Biochemistry, University of California, La Jolla, CA 92093-0314; cInstitute for Microbiology, Ernst-Moritz-Arndt-University of Greifswald, D-17487 Greifswald, Germany; dCenter for Structural Biology, Wake Forest University School of Medicine, Winston-Salem, NC 27157; and eDepartment of Biology, California State University, Fresno, CA 93740 Edited* by Richard M. Losick, Harvard University, Cambridge, MA, and approved February 25, 2010 (received for review January 26, 2010) Bacillithiol (BSH), the α-anomeric glycoside of L-cysteinyl-D-glucosamine disulfide oxidoreductases (TDORs) including thioredoxins (Trx) with L-malic acid, is a major low-molecular-weight thiol in Bacillus sub- and glutaredoxins (5). Oxidized glutaredoxins are reduced by GSH tilis and related bacteria. Here, we identify genes required for BSH generating oxidized GSSG, which is in turn reduced by glutathione biosynthesis and provide evidence that the synthetic pathway has sim- reductase at the expense of NADPH. Thioredoxins are directly ilarities to that established for the related thiol (mycothiol) in the reduced by Trx reductase. Actinobacteria. Consistent with a key role for BSH in detoxification of In addition to disulfide bond formation, thiols can function as electrophiles, the BshA glycosyltransferase and BshB1 deacetylase are nucleophiles to form S-conjugates (6, 7). Thiols react rapidly with encoded in an operon with methylglyoxal synthase. BshB1 is partially some alkylating agents (e.g., N-ethylmaleimide, iodoacetamide) redundant in function with BshB2, a deacetylase of the LmbE family. -
Investigations Into Intracellular Thiols of Biological Importance
Investigations into Intracellular Thiols of Biological Importance by Christine Elizabeth Hand A thesis presented to the University of Waterloo in fulfillment of the thesis requirement for the degree of Doctor of Philosophy in Chemistry Waterloo, Ontario, Canada, 2007 © Christine Elizabeth Hand 2007 AUTHOR'S DECLARATION I hereby declare that I am the sole author of this thesis. This is a true copy of the thesis, including any required final revisions, as accepted by my examiners. I understand that my thesis may be made electronically available to the public. ii Abstract The presence of thiols in living systems is critical for the maintenance of cellular redox homeostasis, the maintenance of protein thiol-disulfide ratios and the protection of cells from reactive oxygen species. In addition to the well studied tripeptide glutathione (γ-Glu-Cys-Gly), a number of compounds have been identified that contribute to these essential cellular roles. Many of these molecules are of great clinical interest due to their essential role in the biochemistry of a number of deadly pathogens, as well as their possible role as therapeutic agents in the treatment of a number of diseases. A series of studies were undertaken using theoretical, chemical and biochemical approaches on a selection of thiols, ergothioneine, the ovothiols and mycothiol, to further our understanding of these necessary biological components. Ergothioneine is present at significant physiological levels in humans and other mammals; however, a definitive role for this thiol has yet to be determined. It has been implicated in radical scavenging in vivo and shows promise as a therapeutic agent against disease states caused by oxidative damage. -
Structure and Functional Diversity of GCN5-Related N-Acetyltransferases (GNAT)
International Journal of Molecular Sciences Review Structure and Functional Diversity of GCN5-Related N-Acetyltransferases (GNAT) Abu Iftiaf Md Salah Ud-Din 1, Alexandra Tikhomirova 1 and Anna Roujeinikova 1,2,* 1 Infection and Immunity Program, Monash Biomedicine Discovery Institute; Department of Microbiology, Monash University, Clayton, Victoria 3800, Australia; [email protected] (A.I.M.S.U.-D.); [email protected] (A.T.) 2 Department of Biochemistry and Molecular Biology, Monash University, Clayton, Victoria 3800, Australia * Correspondence: [email protected]; Tel.: +61-3-9902-9194; Fax: +61-3-9902-9222 Academic Editor: Claudiu T. Supuran Received: 30 May 2016; Accepted: 20 June 2016; Published: 28 June 2016 Abstract: General control non-repressible 5 (GCN5)-related N-acetyltransferases (GNAT) catalyze the transfer of an acyl moiety from acyl coenzyme A (acyl-CoA) to a diverse group of substrates and are widely distributed in all domains of life. This review of the currently available data acquired on GNAT enzymes by a combination of structural, mutagenesis and kinetic methods summarizes the key similarities and differences between several distinctly different families within the GNAT superfamily, with an emphasis on the mechanistic insights obtained from the analysis of the complexes with substrates or inhibitors. It discusses the structural basis for the common acetyltransferase mechanism, outlines the factors important for the substrate recognition, and describes the mechanism of action of inhibitors of these enzymes. It is anticipated that understanding of the structural basis behind the reaction and substrate specificity of the enzymes from this superfamily can be exploited in the development of novel therapeutics to treat human diseases and combat emerging multidrug-resistant microbial infections. -
Report on “Thiol Levels in Bacillus Species
Report on “Thiol levels in Bacillus species exposed to ultraviolet radiation” (NASA Astrobiology Program - Minority Institution Support Faculty Research Awards: August 22, 2011- May 21, 2012) Submitted November 25, 2012 As mandated by the Committee of Space Research, space-faring nations must take precautions in preventing contamination of extraterrestrial bodies by limiting the amount of microbes present to the greatest possible extent. Surveys of spacecraft assembly clean rooms for microbes have revealed the existence of strains of bacteria resistant to high levels of ultraviolet (UV) radiation and vaporous hydrogen peroxide, which are used to sterilize clean rooms. In order to develop effective ways to eradicate these bacteria prior to the spacecraft leaving Earth, we must understand how these bacteria are able to survive these extremophilic conditions. We are particularly concerned about spore forming bacteria, such as Bacillus pumilus SAFR-032 and Bacillus horneckaie, which are highly resistant to UV and oxidative stress. First, we have demonstrated that these species like other Bacilli contains a novel low molecular weight thiol (LMW), bacillithiol. LMW thiols like bacillithiol play a critical role in maintaining a reducing environment and are involved in protection of organisms against a variety of stresses. Bacillithiol has been shown to protect against hypochlorite stress by S-bacillithiolation of cysteines in critical proteins such as glyceraldehyde- 3-phosphate dehydrogenase. We have examined samples of B. pumilus SAFR-032 spores exposed to four different extreme conditions at the International Space Center: (1) deep space, (2) Martian atmosphere, (3) deep space with UV radiation, and (4) Martian atmosphere with UV radiation. Thiol analysis of the surviving spores indicates that levels of bacillithiol are ten times higher in UV radiation treated samples exposed to both deep space and Martian atmosphere conditions. -
Catalysis of Peroxide Reduction by Fast Reacting Protein Thiols Focus Review †,‡ †,‡ ‡,§ ‡,§ ∥ Ari Zeida, Madia Trujillo, Gerardo Ferrer-Sueta, Ana Denicola, Darío A
Review Cite This: Chem. Rev. 2019, 119, 10829−10855 pubs.acs.org/CR Catalysis of Peroxide Reduction by Fast Reacting Protein Thiols Focus Review †,‡ †,‡ ‡,§ ‡,§ ∥ Ari Zeida, Madia Trujillo, Gerardo Ferrer-Sueta, Ana Denicola, Darío A. Estrin, and Rafael Radi*,†,‡ † ‡ § Departamento de Bioquímica, Centro de Investigaciones Biomedicaś (CEINBIO), Facultad de Medicina, and Laboratorio de Fisicoquímica Biologica,́ Facultad de Ciencias, Universidad de la Republica,́ 11800 Montevideo, Uruguay ∥ Departamento de Química Inorganica,́ Analítica y Química-Física and INQUIMAE-CONICET, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, 2160 Buenos Aires, Argentina ABSTRACT: Life on Earth evolved in the presence of hydrogen peroxide, and other peroxides also emerged before and with the rise of aerobic metabolism. They were considered only as toxic byproducts for many years. Nowadays, peroxides are also regarded as metabolic products that play essential physiological cellular roles. Organisms have developed efficient mechanisms to metabolize peroxides, mostly based on two kinds of redox chemistry, catalases/peroxidases that depend on the heme prosthetic group to afford peroxide reduction and thiol-based peroxidases that support their redox activities on specialized fast reacting cysteine/selenocysteine (Cys/Sec) residues. Among the last group, glutathione peroxidases (GPxs) and peroxiredoxins (Prxs) are the most widespread and abundant families, and they are the leitmotif of this review. After presenting the properties and roles of different peroxides in biology, we discuss the chemical mechanisms of peroxide reduction by low molecular weight thiols, Prxs, GPxs, and other thiol-based peroxidases. Special attention is paid to the catalytic properties of Prxs and also to the importance and comparative outlook of the properties of Sec and its role in GPxs. -
5. RPA161724558016.Pdf
93 | P a g e International Standard Serial Number (ISSN): 2319-8141 International Journal of Universal Pharmacy and Bio Sciences 6(6): November-December 2017 INTERNATIONAL JOURNAL OF UNIVERSAL PHARMACY AND BIO SCIENCES IMPACT FACTOR 4.018*** ICV 6.16*** Pharmaceutical Sciences REVIEW ARTICLE …………!!! “GENETICS OF MYCOBACTERIA AND ITS TRENDS IN TUBERCULOSIS: A SYSTEMATIC REVIEW” Mr. Shaikh Zuber Peermohammed*, Dr. Bhise Satish Balkrishna, Sinhgad Technical Education Society‘s (STES – SINHGAD INSTITUTE) Smt. Kashibai Navale College of Pharmacy, Kondhwa, Pune – 411048 (MS) Affiliated to Savitribai Phule Pune University (Formerly known as University of Pune.). KEYWORDS: ABSTRACT Tuberculosis has reemerged as a serious threat to human health because M.tuberculosis, of the increasing prevalence of drugresistant strains and synergetic Lipoarabinomannan, infection with HIV, prompting an urgent need for new and more efficient Mycothiol (MSH), One- treatments. Mycobacterium tuberculosis (M.tb.), an intracellular Hybrid System Vector, pathogen, is exquisitely adapted for human parasitization. Mycobacterial Ubiquitination. infection has been a major cause of death throughout human history and For Correspondence: still results in the death of about two million people globally each year. Mr. Shaikh Zuber This enduring pathogenicity suggests that Mycobacterium may use Peermohammed* unique pathogenic mechanisms during its infection process. M.tb. Address: Department of confronts a more hostile environment during infection, including Pharmacology (Doctoral restricted access to nutrients and reduced oxygen tension. Therefore, Section), Sinhgad M.tb. must possess genetic mechanisms to facilitate integrated responses Technical Education to multiple stresses encountered within the phagosome, and also to Society‘s Smt. Kashibai trigger some yet-to-be-identified switches during infection process. -
Nanaomycin I and J: New Nanaomycins Generated by Mycothiol-Mediated Compounds from “Streptomyces Rosa Subsp
Journal of Bioscience and Bioengineering VOL. 127 No. 5, 549e553, 2019 www.elsevier.com/locate/jbiosc Nanaomycin I and J: New nanaomycins generated by mycothiol-mediated compounds from “Streptomyces rosa subsp. notoensis” OS-3966 Hirotaka Matsuo,1,2 Yoshihiko Noguchi,1,2 Akira Také,3 Jun Nakanishi,4 Katsumi Shigemura,5 Toshiaki Sunazuka,1,2 Yoko Takahashi,1 Satoshi Omura,1 and Takuji Nakashima1,2,* Kitasato Institute for Life Sciences, Kitasato University, 5-9-1 Shirokane, Minato-ku, Tokyo 108-8641, Japan,1 Graduate School of Infection Control Sciences, Kitasato University, 5-9-1 Shirokane, Minato-ku, Tokyo 108-8641, Japan,2 Research Organization for Nano and Life Innovation, Waseda University, 513 Wasedatsurumaki-cho, Shinjuku-ku, Tokyo 162-0041, Japan,3 World Premier International (WPI) Research Center Initiative, International Center for Materials Nanoarchitectonics (MANA), National Institute for Materials Science (NIMS), 1-1 Namiki, Tsukuba, Ibaraki 305-0044, Japan,4 and Department of Urology, Kobe University Graduate School of Medicine, 7-5-2 Kusunokicho, Kobe Chuo-ku, Hyogo 650-0017, Japan5 Received 13 June 2018; accepted 14 October 2018 Available online 29 November 2018 Two new nanaomycin analogs, nanaomycin I and J, were isolated from a cultured broth of an actinomycete strain, “Streptomyces rosa subsp. notoensis” OS-3966. In our previous study, we have confirmed the occurrence of nanaomycin I D (m/z [ 482 [M D H] ) that lacks a pseudo-disaccharide from the mycothiol of nanaomycin H under same culture condition. In this study, to confirm the structure of nanaomycin I, the strain “S. rosa subsp. notoensis” OS-3966 was re- D cultured and the target compound with m/z [ 482 [M D H] was isolated. -
Hatzios Thesis Formatted
Investigations of Metabolic Pathways in Mycobacterium tuberculosis by Stavroula K Hatzios A dissertation submitted in partial satisfaction of the requirements for the degree of Doctor of Philosophy in Chemistry in the Graduate Division of the University of California, Berkeley Committee in charge: Professor Carolyn R. Bertozzi, Chair Professor Matthew B. Francis Professor Tom Alber Fall 2010 Investigations of Metabolic Pathways in Mycobacterium tuberculosis © 2010 By Stavroula K Hatzios Abstract Investigations of Metabolic Pathways in Mycobacterium tuberculosis by Stavroula K Hatzios Doctor of Philosophy in Chemistry University of California, Berkeley Professor Carolyn R. Bertozzi, Chair Mycobacterium tuberculosis (Mtb), the bacterium that causes tuberculosis in humans, infects roughly two billion people worldwide. However, less than one percent of infected individuals are symptomatic. Most have a latent infection characterized by dormant, non- replicating bacteria that persist within a mass of immune cells in the lung called the granuloma. The granuloma provides a protective barrier between infected cells and surrounding tissue. When host immunity is compromised, the granuloma can deteriorate and reactivate the disease. In order to mount a latent infection, Mtb must survive in alveolar macrophages, the host’s primary line of defense against this intracellular pathogen. By evading typical bactericidal processes, Mtb is able to replicate and stimulate granuloma formation. The mechanisms by which Mtb persists in macrophages are ill defined; thus, elucidating the factors responsible for this hallmark of Mtb pathogenesis is an important area of research. This thesis explores three discrete metabolic pathways in Mtb that are likely to mediate its interactions with host immune cells. The first three chapters examine the sulfate assimilation pathway of Mtb and its regulation by the phosphatase CysQ. -
Homocysteine Editing, Thioester Chemistry, Coenzyme A, and the Origin of Coded Peptide Synthesis †
Review Homocysteine Editing, Thioester Chemistry, Coenzyme A, and the Origin of Coded † Peptide Synthesis Hieronim Jakubowski 1,2,* 1 Department of Microbiology, Biochemistry and Molecular Genetics, New Jersey Medical School, Rutgers University, Newark, NJ 07103, USA 2 Department of Biochemistry and Biotechnology, University of Life Sciences, Poznan 60-632, Poland * Correspondence: [email protected] or [email protected]; Tel.: +1-973-972-8733 † Presented at the Banbury Center, Cold Spring Harbor Laboratory, NY meeting on “Evolution of the Translational Apparatus and implication for the origin of the Genetic Code”, 13–16 November 2016. Academic Editor: Koji Tamura Received: 03 January 2017; Accepted: 03 February 2017; Published: 09 February 2017 Abstract: Aminoacyl-tRNA synthetases (AARSs) have evolved “quality control” mechanisms which prevent tRNA aminoacylation with non-protein amino acids, such as homocysteine, homoserine, and ornithine, and thus their access to the Genetic Code. Of the ten AARSs that possess editing function, five edit homocysteine: Class I MetRS, ValRS, IleRS, LeuRS, and Class II LysRS. Studies of their editing function reveal that catalytic modules of these AARSs have a thiol-binding site that confers the ability to catalyze the aminoacylation of coenzyme A, pantetheine, and other thiols. Other AARSs also catalyze aminoacyl-thioester synthesis. Amino acid selectivity of AARSs in the aminoacyl thioesters formation reaction is relaxed, characteristic of primitive amino acid activation systems that may have originated in the Thioester World. With homocysteine and cysteine as thiol substrates, AARSs support peptide bond synthesis. Evolutionary origin of these activities is revealed by genomic comparisons, which show that AARSs are structurally related to proteins involved in coenzyme A/sulfur metabolism and non-coded peptide bond synthesis. -
Ysu1347888245.Pdf (2.08
Chemical and Biochemical Studies of Bacillithiol by Janelle P. N. Russell Submitted in Partial Fulfillment of the Requirements for the Degree of Master of Science in the Chemistry Program YOUNGSTOWN STATE UNIVERSITY August, 2012 Chemical and Biochemical Studies of Bacillithiol Janelle P. N. Russell I hereby release this thesis to the public. I understand that this thesis will be made available from the OhioLINK ETD Center and the Maag Library Circulation Desk for public access. I also authorize the University or other individuals to make copies of this thesis as needed for scholarly research. Signature: Janelle P. N. Russell, Student Date Approvals: Dr. Nina V. Stourman, Thesis Advisor Date Dr. Peter Norris, Committee Member Date Dr. Michael A. Serra, Committee Member Date Peter J. Kasvinsky, Dean of School of Graduate Studies and Research Date Abstract Bacillithiol (BSH), the α-anomeric glycoside of L-cysteinyl-D-glucosamine with L-malic acid, is the recently identified low-molecular-weight (LMW) thiol in low G + C content Gram-positive bacteria. Although BSH has not been fully characterized it is thought to have functions analogous to those of glutathione, the major LMW thiol present in eukaryotes. Glutathione is known to act as an antioxidant and functions to protect the cell against oxidation and other toxins. BSH has been shown to contribute to the detoxification of toxic compounds including some antibiotics. Since bacillithiol is not present in humans its metabolism presents novel targets for antibacterial drug therapies. An attempt was made to chemically synthesize BSH for its use in biochemical studies that would further elucidate its functions.