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Biochemical and molecular characterization of N66 from the shell of Pinctada mazatlanica.

Authors
  • Rivera-Perez, Crisalejandra1
  • Magallanes-Dominguez, Catalina2
  • Dominguez-Beltran, Rosa Virginia3
  • Ojeda-Ramirez de Areyano, Josafat Jehu2
  • Hernandez-Saavedra, Norma Y2
  • 1 Department of Fisheries Ecology, CONACyT-Centro de Investigaciones Biologicas del Noroeste (CIBNOR), La Paz, Baja California Sur, Mexico. , (Mexico)
  • 2 Department of Fisheries Ecology, Molecular Genetics Laboratory, Centro de Investigaciones Biologicas del Noroeste (CIBNOR), La Paz, Baja California Sur, Mexico. , (Mexico)
  • 3 Tecnológico Nacional de México, La Paz, Baja California Sur, Mexico. , (Mexico)
Type
Published Article
Journal
PeerJ
Publisher
PeerJ
Publication Date
Jan 01, 2019
Volume
7
Identifiers
DOI: 10.7717/peerj.7212
PMID: 31293836
Source
Medline
Keywords
Language
English
License
Unknown

Abstract

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.

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