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

AbstractBiomineralization is a widespread biological process, involved in the formation of shells, teeth, and bones. Shell matrix proteins have been widely studied for their importance during shell formation. In 2015, our group identified 72 unique shell matrix proteins in Pinctada fucata, among which PU14 is a matrix protein detected in the soluble fraction that solely exists in the prismatic layer. However, the function of PU14 is still unclear. In this study, the full-length cDNA sequence of PU14 was obtained and functional analyses of PU14 protein during shell formation were performed. The deduced protein has a molecular mass of 77.8 kDa and an isoelectric point of 11.34. The primary protein structure contains Gln-rich and random repeat units, which are typical characteristics of matrix protein and indicate its potential function during shell formation. In vivo and in vitro experiments indicated PU14 has prismatic layer functions during shell formation. The tissue expression patterns showed that PU14 was mainly expressed in the mantle tissue, which is consistent with prismatic layer formation. Notching experiments suggested that PU14 responded to repair and regenerate the injured shell. After inhibiting gene expression by injecting PU14-specific double-stranded RNA, the inner surface of the prismatic layer changed significantly and became rougher. Further, in vitro experiments showed that recombinant protein rPU14 impacted calcite crystal morphology. Taken together, characterization and functional analyses of a novel matrix protein, PU14, provide new insights about basic matrix proteins and their functions during shell formation.

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Recent Advances of Shell Matrix Proteins and Cellular Orchestration in Marine Molluscan Shell Biomineralization

Frontiers in Marine Science ◽

10.3389/fmars.2019.00041 ◽

2019 ◽

Vol 6 ◽

Cited By ~ 10

Author(s):

Xiaorui Song ◽

Zhaoqun Liu ◽

Lingling Wang ◽

Linsheng Song

Keyword(s):

Matrix Proteins ◽

Molluscan Shell ◽

Recent Advances ◽

Shell Matrix ◽

Shell Matrix Proteins

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Shell matrix proteins of the clam, Mya truncata: Roles beyond shell formation through proteomic study

Marine Genomics ◽

10.1016/j.margen.2016.03.005 ◽

2016 ◽

Vol 27 ◽

pp. 69-74 ◽

Cited By ~ 25

Author(s):

Jaison Arivalagan ◽

Benjamin Marie ◽

Victoria A. Sleight ◽

Melody S. Clark ◽

Sophie Berland ◽

...

Keyword(s):

Matrix Proteins ◽

Shell Formation ◽

Shell Matrix ◽

Proteomic Study ◽

Shell Matrix Proteins

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The role of phosphorylation and dephosphorylation of shell matrix proteins in shell formation: an in vivo and in vitro study

CrystEngComm ◽

10.1039/c8ce00755a ◽

2018 ◽

Vol 20 (27) ◽

pp. 3905-3916 ◽

Cited By ~ 4

Author(s):

Jinzhe Du ◽

Guangrui Xu ◽

Chuang Liu ◽

Rongqing Zhang

Keyword(s):

Calcium Carbonate ◽

In Vitro Study ◽

Matrix Proteins ◽

Shell Formation ◽

Carbonate Formation ◽

Shell Matrix ◽

Shell Matrix Proteins

Phosphorylation of shell matrix proteins is critical for shell formation in vivo and can modulate calcium carbonate formation in vitro.

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Culture of mantle epithelial cells expressing shell matrix proteins from scallop Patinopecten yessoensis

Fisheries Science ◽

10.1111/j.1444-2906.2006.01286.x ◽

2006 ◽

Vol 72 (6) ◽

pp. 1277-1285 ◽

Cited By ~ 3

Author(s):

Mayumi ENDOH ◽

Yasushi HASEGAWA

Keyword(s):

Epithelial Cells ◽

Matrix Proteins ◽

Patinopecten Yessoensis ◽

Shell Matrix ◽

Shell Matrix Proteins

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The ‘Shellome’ of the Crocus Clam Tridacna crocea Emphasizes Essential Components of Mollusk Shell Biomineralization

Frontiers in Genetics ◽

10.3389/fgene.2021.674539 ◽

2021 ◽

Vol 12 ◽

Author(s):

Takeshi Takeuchi ◽

Manabu Fujie ◽

Ryo Koyanagi ◽

Laurent Plasseraud ◽

Isabelle Ziegler-Devin ◽

...

Keyword(s):

Organic Matrix ◽

The Other ◽

Matrix Proteins ◽

Shell Formation ◽

Molluscan Shell ◽

Shell Matrix ◽

Essential Components ◽

Shell Matrix Proteins ◽

Shell Organic Matrix ◽

Tridacna Crocea

Molluscan shells are among the most fascinating research objects because of their diverse morphologies and textures. The formation of these delicate biomineralized structures is a matrix-mediated process. A question that arises is what are the essential components required to build these exoskeletons. In order to understand the molecular mechanisms of molluscan shell formation, it is crucial to identify organic macromolecules in different shells from diverse taxa. In the case of bivalves, however, taxon sampling in previous shell proteomics studies are focused predominantly on representatives of the class Pteriomorphia such as pearl oysters, edible oysters and mussels. In this study, we have characterized the shell organic matrix from the crocus clam, Tridacna crocea, (Heterodonta) using various biochemical techniques, including SDS-PAGE, FT-IR, monosaccharide analysis, and enzyme-linked lectin assay (ELLA). Furthermore, we have identified a number of shell matrix proteins (SMPs) using a comprehensive proteomics approach combined to RNA-seq. The biochemical studies confirmed the presence of proteins, polysaccharides, and sulfates in the T. crocea shell organic matrix. Proteomics analysis revealed that the majority of the T. crocea SMPs are novel and dissimilar to known SMPs identified from the other bivalve species. Meanwhile, the SMP repertoire of the crocus clam also includes proteins with conserved functional domains such as chitin-binding domain, VWA domain, and protease inhibitor domain. We also identified BMSP (Blue Mussel Shell Protein, originally reported from Mytilus), which is widely distributed among molluscan shell matrix proteins. Tridacna SMPs also include low-complexity regions (LCRs) that are absent in the other molluscan genomes, indicating that these genes may have evolved in specific lineage. These results highlight the diversity of the organic molecules – in particular proteins – that are essential for molluscan shell formation.

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