Sequence Analysis, Kinetic Constants, and Anion Inhibition Profile of the Nacrein-Like Protein (CgiNAP2X1) from the Pacific Oyster Magallana gigas (Ex-Crassostrea gigas)
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 react...
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Online Access: | http://hdl.handle.net/11588/683350 https://doi.org/10.3390/md15090270 |
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ftunivnapoliiris:oai:www.iris.unina.it:11588/683350 2024-06-23T07:52:16+00:00 Sequence Analysis, Kinetic Constants, and Anion Inhibition Profile of the Nacrein-Like Protein (CgiNAP2X1) from the Pacific Oyster Magallana gigas (Ex-Crassostrea gigas) Perfetto, Rosa SANSONE, GIOVANNI BARONE, CARMELA MARIA ASSUNTA ROSSI, MOSE' CAPASSO, CLEMENTE Del Prete, Sonia Vullo, Daniela Supuran, Claudiu T Perfetto, Rosa Del Prete, Sonia Vullo, Daniela Sansone, Giovanni Barone, CARMELA MARIA ASSUNTA Rossi, Mose' Supuran, Claudiu T Capasso, Clemente 2017 http://hdl.handle.net/11588/683350 https://doi.org/10.3390/md15090270 eng eng info:eu-repo/semantics/altIdentifier/wos/WOS:000411559800009 volume:15 issue:9 firstpage:270 lastpage:284 numberofpages:15 journal:MARINE DRUGS http://hdl.handle.net/11588/683350 doi:10.3390/md15090270 info:eu-repo/semantics/altIdentifier/scopus/2-s2.0-85030980541 info:eu-repo/semantics/openAccess anion inhibitor carbonic anhydrase kinetic constant molluskan nacrein-like protein phylogeny signal peptide info:eu-repo/semantics/article 2017 ftunivnapoliiris https://doi.org/10.3390/md15090270 2024-06-10T14:58:50Z 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. Article in Journal/Newspaper Crassostrea gigas Pacific oyster IRIS Università degli Studi di Napoli Federico II Pacific Marine Drugs 15 9 270 |
institution |
Open Polar |
collection |
IRIS Università degli Studi di Napoli Federico II |
op_collection_id |
ftunivnapoliiris |
language |
English |
topic |
anion inhibitor carbonic anhydrase kinetic constant molluskan nacrein-like protein phylogeny signal peptide |
spellingShingle |
anion inhibitor carbonic anhydrase kinetic constant molluskan nacrein-like protein phylogeny signal peptide Perfetto, Rosa SANSONE, GIOVANNI BARONE, CARMELA MARIA ASSUNTA ROSSI, MOSE' CAPASSO, CLEMENTE Del Prete, Sonia Vullo, Daniela Supuran, Claudiu T Sequence Analysis, Kinetic Constants, and Anion Inhibition Profile of the Nacrein-Like Protein (CgiNAP2X1) from the Pacific Oyster Magallana gigas (Ex-Crassostrea gigas) |
topic_facet |
anion inhibitor carbonic anhydrase kinetic constant molluskan nacrein-like protein phylogeny signal peptide |
description |
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. |
author2 |
Perfetto, Rosa Del Prete, Sonia Vullo, Daniela Sansone, Giovanni Barone, CARMELA MARIA ASSUNTA Rossi, Mose' Supuran, Claudiu T Capasso, Clemente |
format |
Article in Journal/Newspaper |
author |
Perfetto, Rosa SANSONE, GIOVANNI BARONE, CARMELA MARIA ASSUNTA ROSSI, MOSE' CAPASSO, CLEMENTE Del Prete, Sonia Vullo, Daniela Supuran, Claudiu T |
author_facet |
Perfetto, Rosa SANSONE, GIOVANNI BARONE, CARMELA MARIA ASSUNTA ROSSI, MOSE' CAPASSO, CLEMENTE Del Prete, Sonia Vullo, Daniela Supuran, Claudiu T |
author_sort |
Perfetto, Rosa |
title |
Sequence Analysis, Kinetic Constants, and Anion Inhibition Profile of the Nacrein-Like Protein (CgiNAP2X1) from the Pacific Oyster Magallana gigas (Ex-Crassostrea gigas) |
title_short |
Sequence Analysis, Kinetic Constants, and Anion Inhibition Profile of the Nacrein-Like Protein (CgiNAP2X1) from the Pacific Oyster Magallana gigas (Ex-Crassostrea gigas) |
title_full |
Sequence Analysis, Kinetic Constants, and Anion Inhibition Profile of the Nacrein-Like Protein (CgiNAP2X1) from the Pacific Oyster Magallana gigas (Ex-Crassostrea gigas) |
title_fullStr |
Sequence Analysis, Kinetic Constants, and Anion Inhibition Profile of the Nacrein-Like Protein (CgiNAP2X1) from the Pacific Oyster Magallana gigas (Ex-Crassostrea gigas) |
title_full_unstemmed |
Sequence Analysis, Kinetic Constants, and Anion Inhibition Profile of the Nacrein-Like Protein (CgiNAP2X1) from the Pacific Oyster Magallana gigas (Ex-Crassostrea gigas) |
title_sort |
sequence analysis, kinetic constants, and anion inhibition profile of the nacrein-like protein (cginap2x1) from the pacific oyster magallana gigas (ex-crassostrea gigas) |
publishDate |
2017 |
url |
http://hdl.handle.net/11588/683350 https://doi.org/10.3390/md15090270 |
geographic |
Pacific |
geographic_facet |
Pacific |
genre |
Crassostrea gigas Pacific oyster |
genre_facet |
Crassostrea gigas Pacific oyster |
op_relation |
info:eu-repo/semantics/altIdentifier/wos/WOS:000411559800009 volume:15 issue:9 firstpage:270 lastpage:284 numberofpages:15 journal:MARINE DRUGS http://hdl.handle.net/11588/683350 doi:10.3390/md15090270 info:eu-repo/semantics/altIdentifier/scopus/2-s2.0-85030980541 |
op_rights |
info:eu-repo/semantics/openAccess |
op_doi |
https://doi.org/10.3390/md15090270 |
container_title |
Marine Drugs |
container_volume |
15 |
container_issue |
9 |
container_start_page |
270 |
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1802643522224914432 |