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In-depth proteomic analysis of shell matrix proteins of Pinctada fucata.

Liu C, Li S, Kong J, Liu Y, Wang T, Xie L, Zhang R - Sci Rep (2015)

Bottom Line: Identification of diverse SMPs will lay a foundation for understanding biomineralization process.Immunohistological localization techniques identify the SMPs in the mantle, shells and synthetic calcite.Together, these proteomic data increase the repertoires of the shell matrix proteins in P. fucata and suggest that shell formation in P. fucata may involve tight regulation of cellular activities and the extracellular microenvironment.

View Article: PubMed Central - PubMed

Affiliation: Institute of Marine Biotechnology, Collaborative Innovation Center of Deep Sea Biology, School of Life Sciences, Tsinghua University, Beijing 100084 China.

ABSTRACT
The shells of pearl oysters, Pinctada fucata, are composed of calcite and aragonite and possess remarkable mechanical properties. These shells are formed under the regulation of macromolecules, especially shell matrix proteins (SMPs). Identification of diverse SMPs will lay a foundation for understanding biomineralization process. Here, we identified 72 unique SMPs using liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis of proteins extracted from the shells of P. fucata combined with a draft genome. Of 72 SMPs, 17 SMPs are related to both the prismatic and nacreous layers. Moreover, according to the diverse domains found in the SMPs, we hypothesize that in addition to controlling CaCO3 crystallization and crystal organization, these proteins may potentially regulate the extracellular microenvironment and communicate between cells and the extracellular matrix (ECM). Immunohistological localization techniques identify the SMPs in the mantle, shells and synthetic calcite. Together, these proteomic data increase the repertoires of the shell matrix proteins in P. fucata and suggest that shell formation in P. fucata may involve tight regulation of cellular activities and the extracellular microenvironment.

No MeSH data available.


Related in: MedlinePlus

(a) Optical image shows the prismatic and nacreous layer of a typical shell (red box is examined by SEM), (b) SEM shows the shell surfaces of Pinctada fucata: the prismatic (b) and nacreous layer (c,d) SDS-PAGE of the four groups of extracted proteins (ESM and EISM are EDTA-soluble and EDTA-insoluble extracts, respectively; P and N mean the prismatic and nacreous layer, respectively).
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f1: (a) Optical image shows the prismatic and nacreous layer of a typical shell (red box is examined by SEM), (b) SEM shows the shell surfaces of Pinctada fucata: the prismatic (b) and nacreous layer (c,d) SDS-PAGE of the four groups of extracted proteins (ESM and EISM are EDTA-soluble and EDTA-insoluble extracts, respectively; P and N mean the prismatic and nacreous layer, respectively).

Mentions: The shell of P. fucata is composed of two layers, the prismatic layer and the nacreous layer (Fig. 1a). The prismatic layer is composed of prisms with length of 10–40 μm embedded in the organic sheath (Fig. 1b), and the nacreous layer is formed by stacked hexagonal nanotablets with side lengths of 0.5–3 μm (Fig. 1c). To extract SMPs, shells were first cleaned with NaOH to avoid possible contamination from outside organic matter. Then, separated shells, prism and nacre were dissolved with EDTA, leaving soluble and insoluble extracts. EDTA can chelate Ca2+, dissolve the shell and release the organic matrices. In this study, we observed that after 60 h, the shell can be fully dissolved. The yields of organic matrices from the shell were approximately 1.5–3.5 mg/g (determined by the concentration of proteins obtained from certain amounts of shell powders).


In-depth proteomic analysis of shell matrix proteins of Pinctada fucata.

Liu C, Li S, Kong J, Liu Y, Wang T, Xie L, Zhang R - Sci Rep (2015)

(a) Optical image shows the prismatic and nacreous layer of a typical shell (red box is examined by SEM), (b) SEM shows the shell surfaces of Pinctada fucata: the prismatic (b) and nacreous layer (c,d) SDS-PAGE of the four groups of extracted proteins (ESM and EISM are EDTA-soluble and EDTA-insoluble extracts, respectively; P and N mean the prismatic and nacreous layer, respectively).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4660305&req=5

f1: (a) Optical image shows the prismatic and nacreous layer of a typical shell (red box is examined by SEM), (b) SEM shows the shell surfaces of Pinctada fucata: the prismatic (b) and nacreous layer (c,d) SDS-PAGE of the four groups of extracted proteins (ESM and EISM are EDTA-soluble and EDTA-insoluble extracts, respectively; P and N mean the prismatic and nacreous layer, respectively).
Mentions: The shell of P. fucata is composed of two layers, the prismatic layer and the nacreous layer (Fig. 1a). The prismatic layer is composed of prisms with length of 10–40 μm embedded in the organic sheath (Fig. 1b), and the nacreous layer is formed by stacked hexagonal nanotablets with side lengths of 0.5–3 μm (Fig. 1c). To extract SMPs, shells were first cleaned with NaOH to avoid possible contamination from outside organic matter. Then, separated shells, prism and nacre were dissolved with EDTA, leaving soluble and insoluble extracts. EDTA can chelate Ca2+, dissolve the shell and release the organic matrices. In this study, we observed that after 60 h, the shell can be fully dissolved. The yields of organic matrices from the shell were approximately 1.5–3.5 mg/g (determined by the concentration of proteins obtained from certain amounts of shell powders).

Bottom Line: Identification of diverse SMPs will lay a foundation for understanding biomineralization process.Immunohistological localization techniques identify the SMPs in the mantle, shells and synthetic calcite.Together, these proteomic data increase the repertoires of the shell matrix proteins in P. fucata and suggest that shell formation in P. fucata may involve tight regulation of cellular activities and the extracellular microenvironment.

View Article: PubMed Central - PubMed

Affiliation: Institute of Marine Biotechnology, Collaborative Innovation Center of Deep Sea Biology, School of Life Sciences, Tsinghua University, Beijing 100084 China.

ABSTRACT
The shells of pearl oysters, Pinctada fucata, are composed of calcite and aragonite and possess remarkable mechanical properties. These shells are formed under the regulation of macromolecules, especially shell matrix proteins (SMPs). Identification of diverse SMPs will lay a foundation for understanding biomineralization process. Here, we identified 72 unique SMPs using liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis of proteins extracted from the shells of P. fucata combined with a draft genome. Of 72 SMPs, 17 SMPs are related to both the prismatic and nacreous layers. Moreover, according to the diverse domains found in the SMPs, we hypothesize that in addition to controlling CaCO3 crystallization and crystal organization, these proteins may potentially regulate the extracellular microenvironment and communicate between cells and the extracellular matrix (ECM). Immunohistological localization techniques identify the SMPs in the mantle, shells and synthetic calcite. Together, these proteomic data increase the repertoires of the shell matrix proteins in P. fucata and suggest that shell formation in P. fucata may involve tight regulation of cellular activities and the extracellular microenvironment.

No MeSH data available.


Related in: MedlinePlus