Abstract
Biomineralization, including shell formation, is dedicatedly regulated by matrix proteins. PfY2, a matrix protein detected in the ethylene diamine tetraacetic acid (EDTA)-soluble fraction from both prismatic layer and nacreous layer, was discovered by our group using microarray. It may play dual roles during biomineralization. However, the molecular mechanism is still unclear. In this research, we studied the function of PfY2 on crystallization in vivo and in vitro, revealing that it might be a negative regulator during shell formation. Notching experiment indicated that PfY2 was involved in shell repairing and regenerating process. Repression of PfY2 gene affected the structure of prismatic and nacreous layer simultaneously, confirming its dual roles in shell formation. Recombinant protein rPfY2 significantly suppressed CaCO3 precipitation rate, participated in the crystal nucleation process, changed the morphology of crystals and inhibited the transformation of amorphous calcium carbonate (ACC) to stable calcite or aragonite in vitro. Our results may provide new evidence on the biomineralization inhibition process.
Highlights
50 different kinds of shell matrix proteins have been identified and reported until now[11]
The polymerase chain reaction (PCR) and rapid amplification of cDNA ends (RACE) techniques were used to screen new matrix protein members based on available protein sequences
Our global gene expression analysis profiles during larval development showed that most genes involved in biomineralization, including nacrein, pearlin, Pif, ACCBP, Prisilkin-39, Shematrin and KRMP family, were greatly up-regulated in juveniles and maintained a relative high expression level[17]
Summary
50 different kinds of shell matrix proteins have been identified and reported until now[11]. Lysine (K)-rich matrix protein (KRMP) family and Prisilkin-39 were confirmed via these molecular biology and biochemistry measures mentioned above[23, 24]. They are typical cases with the deficiency that only proteins similar to previously identified ones could be found. Prodissoconch II with a homogeneous structure of aragonite and a thin structure of calcite appears at the lateral D-shaped stage or umbonal stage It is not until the juvenile stage has P. fucata formed the comprehensive prismatic and nacreous layers[26]. Those results may provide us deeper insights into the control of shell formation process and the perspective of the synthesis of artificial nacre
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