Sequence Analysis, Kinetic Constants, and Anion Inhibition Profile of the Nacrein-Like Protein (CgiNAP2X1) from the Pacific Oyster Magallana gigas (Ex-Crassostrea gigas)
| Autor: | Sonia Del Prete, Clemente Capasso, Giovanni Sansone, Claudiu T. Supuran, Rosa Perfetto, Daniela Vullo, C.M.A. Barone, Mosè Rossi |
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| Přispěvatelé: | Perfetto, Rosa, Del Prete, Sonia, Vullo, Daniela, Sansone, Giovanni, Barone, CARMELA MARIA ASSUNTA, Rossi, Mose', Supuran, Claudiu T, Capasso, Clemente |
| Jazyk: | angličtina |
| Rok vydání: | 2017 |
| Předmět: |
Anions
Bicarbonate carbonic anhydrase Pharmaceutical Science anion inhibitors anion inhibitor phylogeny 01 natural sciences Article chemistry.chemical_compound Carbonic anhydrase Drug Discovery Animals nacrein-like protein signal peptide Enzyme kinetics Phenylboronic acid molluskan kinetic constants lcsh:QH301-705.5 Pharmacology Toxicology and Pharmaceutics (miscellaneous) Peptide sequence Sulfamide Carbonic Anhydrases chemistry.chemical_classification Pacific Ocean biology 010405 organic chemistry kinetic constant Phenylarsonic acid Ostreidae 0104 chemical sciences Kinetics 010404 medicinal & biomolecular chemistry Enzyme lcsh:Biology (General) chemistry Biochemistry Mollusca biology.protein |
| Zdroj: | Marine drugs (2017). doi:10.3390/md15090270 info:cnr-pdr/source/autori:Rosa Perfetto, Sonia Del Prete, Daniela Vullo, Giovanni Sansone, Carmela M. A. Barone Mosè Rossi, Claudiu T. Supuran * and Clemente Capasso/titolo:Sequence Analysis, Kinetic Constants, and Anion Inhibition Profile of the Nacrein-Like Protein (CgiNAP2X1) from the Pacific Oyster Magallana gigas (Ex-Crassostre.../doi:10.3390%2Fmd15090270/rivista:Marine drugs/anno:2017/pagina_da:/pagina_a:/intervallo_pagine:/volume Marine Drugs Marine Drugs; Volume 15; Issue 9; Pages: 270 Marine Drugs, Vol 15, Iss 9, p 270 (2017) |
| Popis: | The carbonic anhydrase (CA, EC 4.2.1.1) superfamily of metalloenzymes catalyzes the hydration of carbon dioxide to bicarbonate and protons. The catalytically active form of these enzymes incorporates a metal hydroxide derivative, the formation of which is the rate-determining step of catalytic reaction, being affected by the transfer of a proton from a metal-coordinated water molecule to the environment. Here, we report the cloning, expression, and purification of a particular CA, i.e., nacrein-like protein encoded in the genome of the Pacific oyster Magallana gigas (previously known as Crassostrea gigas). Furthermore, the amino acid sequence, kinetic constants, and anion inhibition profile of the recombinant enzyme were investigated for the first time. The new protein, CgiNAP2X1, is highly effective as catalyst for the CO₂ hydration reaction, based on the measured kinetic parameters, i.e., kcat = 1.0 × 10⁶ s(-1) and kcat/KM = 1.2 × 10⁸ M(-1)·s(-1). CgiNAP2X1 has a putative signal peptide, which probably allows an extracellular localization of the protein. The inhibition data demonstrated that the best anion inhibitors of CgiNAP2X1 were diethyldithiocarbamate, sulfamide, sulfamate, phenylboronic acid and phenylarsonic acid, which showed a micromolar affinity for this enzyme, with KIs in the range of 76-87 μM. These studies may add new information on the physiological role of the molluskan CAs in the biocalcification processes. |
| Databáze: | OpenAIRE |
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