Molecular cloning,structure, and reactivity of the second bromoperoxidase from Ascophyllum nodosum |
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| Affiliation: | 1. Fachbereich Chemie, Organische Chemie, Technische Universität Kaiserslautern, Erwin-Schrödinger-Straße, D-67663 Kaiserslautern, Germany;2. Zurmaiener Straße 16, D-54292 Trier, Germany;3. Landesamt für Verbraucherschutz Sachsen-Anhalt, Fachbereich 3 Lebensmittelsicherheit, Postfach 200857, D-06009 Halle (Saale), Germany;4. Helmholtz-Institut für Pharmazeutische Forschung, Helmholtz-Zentrum für Infektionsforschung und Pharmazeutische Biotechnologie, Universität des Saarlandes, Campus, D-66123 Saarbrücken, Germany;5. UPMC Université Paris 6, FR 2424, Station Biologique, F-29680 Roscoff, France;6. CNRS, FR 2424, Station Biologique, F-29680 Roscoff, France;7. UPMC Université Paris 6, UMR 7139 Végétaux marins et Biomolécules, Station Biologique, F-29680 Roscoff, France;8. CNRS, UMR 7139 Végétaux marins et Biomolécules, Station Biologique, F-29680 Roscoff, France;1. ImmunoTechnology Section, Vaccine Research Center, National Institute of Allergy and Infectious Diseases, Bethesda, MD 20892, United States;2. Duke Human Vaccine Institute, Duke University Medical Center, Durham, NC 27710, United States;1. School of Chemical Engineering, Northeast Electric Power University, Jilin, Jilin 132012, China;2. Institute of Electronic Paper Displays, South China Academy of Advanced Optoelectronics, South China Normal University, Guangzhou 510006, China;3. Key Laboratory of Bio-Inspired Smart Interfacial Science and Technology of Ministry of Education, School of Chemistry, Beihang University, Beijing 100191, China;4. Department of Chemistry, Faculty of Science and Arts and Promising Centre for Sensors and Electronic Devices, Najran University, Najran 11001, Saudi Arabia;1. Animal Model Research Center, Jeonbuk Department of Inhalation Research, Korea Institute of Toxicology, Jeongup, Republic of Korea;2. School of Mechanical Engineering, Gwangju Institute of Science and Technology, Gwangju, Republic of Korea;3. Department of Biomedical Science and Engineering, Gwangju Institute of Science and Technology, Gwangju, Republic of Korea;4. Departement of Neurosurgery, Presbyterian Medical Center, Jeonju, Republic of Korea;1. CCMAR, CIMAR-Laboratório Associado, Universidade do Algarve Gambelas, 8005-139 Faro, Portugal;2. Departamento de Ciencias del Mar y Biología Aplicada, Universidad de Alicante, Campus de San Vicente de Raspeig, Ap. 99, E-03080 Alicante, Spain;1. South Texas Center for Emerging Infectious Disease and the Center for Excellence in Infection Genomics, University of Texas at San Antonio, San Antonio, TX 78249, United States;2. Department of Microbiology and Immunology, University of Texas Health Science Center at San Antonio, San Antonio, TX 78229, United States |
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| Abstract: | The sequence of bromoperoxidase II from the brown alga Ascophyllum nodosum was determined from a full length cloned cDNA, obtained from a tandem mass spectrometry RT-PCR-approach. The clone encodes a protein composed of 641 amino-acids, which provides a mature 67.4 kDa-bromoperoxidase II-protein (620 amino-acids). Based on 43% sequence homology with the previously characterized bromoperoxidase I from A. nodosum, a tertiary structure was modeled for the bromoperoxidase II. The structural model was refined on the basis of results from gel filtration and vanadate-binding studies, showing that the bromoperoxidase II is a hexameric metalloprotein, which binds 0.5 equivalents of vanadate as cofactor per 67.4 kDa-subunit, for catalyzing oxidation of bromide by hydrogen peroxide in a bi-bi-ping-pong mechanism (kcat = 153 s−1, 22 °C, pH 5.9). Bromide thereby is converted into a bromoelectrophile of reactivity similar to molecular bromine, based on competition kinetic data on phenol bromination and correlation analysis. Reactivity provided by the bromoperoxidase II mimics biosynthesis of methyl 4-bromopyrrole-2-carboxylate, a natural product isolated from the marine sponge Axinella tenuidigitata. |
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