Cloning, purification, kinetic and anion inhibition studies of a recombinant β-carbonic anhydrase from the Atlantic salmon parasite platyhelminth Gyrodactylus salaris

• Published in: Journal of enzyme inhibition and medicinal chemistry Vol. 37; no. 1; pp. 1577 - 1586
• Main Authors: Aspatwar, Ashok, Barker, Harlan, Aisala, Heidi, Zueva, Ksenia, Kuuslahti, Marianne, Tolvanen, Martti, Primmer, Craig R., Lumme, Jaakko, Bonardi, Alessandro, Tripathi, Amit, Parkkila, Seppo, Supuran, Claudiu T.
• Format: Journal Article
• Language: English
• Published: England Taylor & Francis 01-12-2022; Taylor & Francis Ltd; Taylor & Francis Group
• Subjects: Acetazolamide; Animals; anion inhibitors; Anions; Anions - pharmacology; Carbon dioxide; Carbonic anhydrase; Carbonic Anhydrase Inhibitors - pharmacology; Carbonic anhydrases; Carbonic Anhydrases - genetics; Cloning; Cloning, Molecular; Enzymes; Functional foods & nutraceuticals; Genomes; Glycerol; Gyrodactylus salaris; kinetics; Neurosciences; Original; Parasites; Parasites - genetics; Platyhelminths - genetics; Proteins; Salmo salar - genetics; Salmon; Sulfamide; Sulfonamides; sulphamic acid; Therapeutic targets; sulphamic acid; Carbonic anhydrase; Gyrodactylus salaris; kinetics; anion inhibitors
• ISSN: 1475-6366; 1475-6374
• DOI: 10.1080/14756366.2022.2080818

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, CO
2
+ H
2
O ⇋ HCO
3
−
+ H
+
with a k
cat
of 1.1 × 10
5
s
−1
and a k
cat
/K
m
of 7.58 × 10
6
M
−1
× s
−1
. This activity was inhibited by acetazolamide (K
I
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 (K
I
of 81 µM), N,N-diethyldithiocarbamate (K
I
of 67 µM) and sulphamic acid (K
I
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.