Biochemical and molecular characterization of N66 from the shell of Pinctada mazatlanica
Mollusk shell mineralization is a tightly controlled process made by shell matrix proteins (SMPs). However, the study of SMPs has been limited to a few model species. In this study, the N66 mRNA of the pearl oyster Pinctada mazatlanica was cloned and functionally characterized. The full sequence of...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
PeerJ Inc.
2019-06-01
|
| Series: | PeerJ |
| Subjects: | |
| Online Access: | https://peerj.com/articles/7212.pdf |
| _version_ | 1797418527219515392 |
|---|---|
| author | Crisalejandra Rivera-Perez Catalina Magallanes-Dominguez Rosa Virginia Dominguez-Beltran Josafat Jehu Ojeda-Ramirez de Areyano Norma Y. Hernandez-Saavedra |
| author_facet | Crisalejandra Rivera-Perez Catalina Magallanes-Dominguez Rosa Virginia Dominguez-Beltran Josafat Jehu Ojeda-Ramirez de Areyano Norma Y. Hernandez-Saavedra |
| author_sort | Crisalejandra Rivera-Perez |
| collection | DOAJ |
| description | Mollusk shell mineralization is a tightly controlled process made by shell matrix proteins (SMPs). However, the study of SMPs has been limited to a few model species. In this study, the N66 mRNA of the pearl oyster Pinctada mazatlanica was cloned and functionally characterized. The full sequence of the N66 mRNA comprises 1,766 base pairs, and encodes one N66 protein. A sequence analysis revealed that N66 contained two carbonic anhydrase (CA) domains, a NG domain and several glycosylation sites. The sequence showed similarity to the CA VII but also with its homolog protein nacrein. The native N66 protein was isolated from the shell and identified by mass spectrometry, the peptide sequence matched to the nucleotide sequence obtained. Native N66 is a glycoprotein with a molecular mass of 60–66 kDa which displays CA activity and calcium carbonate precipitation ability in presence of different salts. Also, a recombinant form of N66 was produced in Escherichia coli, and functionally characterized. The recombinant N66 displayed higher CA activity and crystallization capability than the native N66, suggesting that the lack of posttranslational modifications in the recombinant N66 might modulate its activity. |
| first_indexed | 2024-03-09T06:34:12Z |
| format | Article |
| id | doaj.art-668836e9c31b40bb98d41072620e9d7e |
| institution | Directory Open Access Journal |
| issn | 2167-8359 |
| language | English |
| last_indexed | 2024-03-09T06:34:12Z |
| publishDate | 2019-06-01 |
| publisher | PeerJ Inc. |
| record_format | Article |
| series | PeerJ |
| spelling | doaj.art-668836e9c31b40bb98d41072620e9d7e2023-12-03T11:00:28ZengPeerJ Inc.PeerJ2167-83592019-06-017e721210.7717/peerj.7212Biochemical and molecular characterization of N66 from the shell of Pinctada mazatlanicaCrisalejandra Rivera-Perez0Catalina Magallanes-Dominguez1Rosa Virginia Dominguez-Beltran2Josafat Jehu Ojeda-Ramirez de Areyano3Norma Y. Hernandez-Saavedra4Department of Fisheries Ecology, CONACyT-Centro de Investigaciones Biologicas del Noroeste (CIBNOR), La Paz, Baja California Sur, MexicoDepartment of Fisheries Ecology, Molecular Genetics Laboratory, Centro de Investigaciones Biologicas del Noroeste (CIBNOR), La Paz, Baja California Sur, MexicoTecnológico Nacional de México, La Paz, Baja California Sur, MexicoDepartment of Fisheries Ecology, Molecular Genetics Laboratory, Centro de Investigaciones Biologicas del Noroeste (CIBNOR), La Paz, Baja California Sur, MexicoDepartment of Fisheries Ecology, Molecular Genetics Laboratory, Centro de Investigaciones Biologicas del Noroeste (CIBNOR), La Paz, Baja California Sur, MexicoMollusk shell mineralization is a tightly controlled process made by shell matrix proteins (SMPs). However, the study of SMPs has been limited to a few model species. In this study, the N66 mRNA of the pearl oyster Pinctada mazatlanica was cloned and functionally characterized. The full sequence of the N66 mRNA comprises 1,766 base pairs, and encodes one N66 protein. A sequence analysis revealed that N66 contained two carbonic anhydrase (CA) domains, a NG domain and several glycosylation sites. The sequence showed similarity to the CA VII but also with its homolog protein nacrein. The native N66 protein was isolated from the shell and identified by mass spectrometry, the peptide sequence matched to the nucleotide sequence obtained. Native N66 is a glycoprotein with a molecular mass of 60–66 kDa which displays CA activity and calcium carbonate precipitation ability in presence of different salts. Also, a recombinant form of N66 was produced in Escherichia coli, and functionally characterized. The recombinant N66 displayed higher CA activity and crystallization capability than the native N66, suggesting that the lack of posttranslational modifications in the recombinant N66 might modulate its activity.https://peerj.com/articles/7212.pdfBiomineralizationN66SMPsMollusks |
| spellingShingle | Crisalejandra Rivera-Perez Catalina Magallanes-Dominguez Rosa Virginia Dominguez-Beltran Josafat Jehu Ojeda-Ramirez de Areyano Norma Y. Hernandez-Saavedra Biochemical and molecular characterization of N66 from the shell of Pinctada mazatlanica PeerJ Biomineralization N66 SMPs Mollusks |
| title | Biochemical and molecular characterization of N66 from the shell of Pinctada mazatlanica |
| title_full | Biochemical and molecular characterization of N66 from the shell of Pinctada mazatlanica |
| title_fullStr | Biochemical and molecular characterization of N66 from the shell of Pinctada mazatlanica |
| title_full_unstemmed | Biochemical and molecular characterization of N66 from the shell of Pinctada mazatlanica |
| title_short | Biochemical and molecular characterization of N66 from the shell of Pinctada mazatlanica |
| title_sort | biochemical and molecular characterization of n66 from the shell of pinctada mazatlanica |
| topic | Biomineralization N66 SMPs Mollusks |
| url | https://peerj.com/articles/7212.pdf |
| work_keys_str_mv | AT crisalejandrariveraperez biochemicalandmolecularcharacterizationofn66fromtheshellofpinctadamazatlanica AT catalinamagallanesdominguez biochemicalandmolecularcharacterizationofn66fromtheshellofpinctadamazatlanica AT rosavirginiadominguezbeltran biochemicalandmolecularcharacterizationofn66fromtheshellofpinctadamazatlanica AT josafatjehuojedaramirezdeareyano biochemicalandmolecularcharacterizationofn66fromtheshellofpinctadamazatlanica AT normayhernandezsaavedra biochemicalandmolecularcharacterizationofn66fromtheshellofpinctadamazatlanica |