@article{62f64bb42fe84d1b80aaff6359fac99d,
title = "Cloning, purification, kinetic and anion inhibition studies of a recombinant β-carbonic anhydrase from the Atlantic salmon parasite platyhelminth Gyrodactylus salaris",
abstract = "A β-class carbonic anhydrase (CA, EC 4.2.1.1) was cloned from the genome of the Monogenean platyhelminth Gyrodactylus salaris, a parasite of Atlantic salmon. The new enzyme, GsaCAβ has a significant catalytic activity for the physiological reaction, CO2 + H2O ⇋ HCO3− + H+ with a kcat of 1.1 × 105 s−1 and a kcat/Km of 7.58 × 106 M−1 × s−1. This activity was inhibited by acetazolamide (KI of 0.46 µM), a sulphonamide in clinical use, as well as by selected inorganic anions and small molecules. Most tested anions inhibited GsaCAβ at millimolar concentrations, but sulfamide (KI of 81 µM), N,N-diethyldithiocarbamate (KI of 67 µM) and sulphamic acid (KI of 6.2 µM) showed a rather efficient inhibitory action. There are currently very few non-toxic agents effective in combating this parasite. GsaCAβ is subsequently proposed as a new drug target for which effective inhibitors can be designed.",
keywords = "anion inhibitors, Carbonic anhydrase, Gyrodactylus salaris, kinetics, sulphamic acid",
author = "Ashok Aspatwar and Harlan Barker and Heidi Aisala and Ksenia Zueva and Marianne Kuuslahti and Martti Tolvanen and Primmer, {Craig R.} and Jaakko Lumme and Alessandro Bonardi and Amit Tripathi and Seppo Parkkila and Supuran, {Claudiu T.}",
note = "Funding Information: A nearly complete GsaCAβ sequence was obtained from transcriptome data produced at the University of Turku. The open reading frame produced a translation of 229 amino acids, 165 of which were supported by genome data from the University of Oulu. A BLAST search at NCBI suggested that the sequence is close to full length, and therefore the existing sequence was only extended by an ATG codon at the beginning of the transcript and a stop codons (TAA TAG) at the end. This sequence of the β-CA was inserted in a pBVboost vector construct for the production of recombinant protein (). The construct was obtained by GeneArt (Invitrogen, Regensburg, Germany). The sequence of the β-CA was modified accordingly to produce the protein in bacterial (Escherichia coli) cells. Funding Information: All operations with 3 D protein structure models and molecular visualisation were performed using ChimeraX (daily build 1.4.dev202202030703), developed by the UCSF Resource for Biocomputing, Visualisation, and Informatics (San Francisco, California, USA), supported in part by the National Institutes of Health. Funding Information: This research was financed by the Italian Ministry for Education and Science (MIUR), grant PRIN: rot. [2017XYBP2R]; Ente Cassa di Risparmio di Firenze (ECRF), grant [CRF2020.1395] (to CTS); Academy of Finland (to SP); Jane & Aatos Erkko Foundation (to SP); Finnish Cultural Foundation (to AA); and Tampere Tuberculosis Foundation (to AA). Publisher Copyright: {\textcopyright} 2022 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.",
year = "2022",
doi = "10.1080/14756366.2022.2080818",
language = "English",
volume = "37",
pages = "1577--1586",
journal = "JOURNAL OF ENZYME INHIBITION AND MEDICINAL CHEMISTRY",
issn = "1475-6366",
publisher = "Informa Healthcare",
number = "1",
}