Identification and Comparison of Amorphous Calcium Carbonate-Binding Protein and Acetylcholine-Binding Protein in the Abalone, Haliotis discus hannai

Nacre has two different microarchitectures: columnar nacre and sheet nacre. We previously identified an important regulator of the morphology of sheet nacre tablets, which was named amorphous calcium carbonate-binding protein (pf-ACCBP). However, little is known about its counterpart in columnar nacre. Moreover, pf-ACCBP shares significant sequence similarity with a group of acetylcholine-binding proteins (AChBP) that participate in neuronal synapses transmission, but the relationships between the two proteins, which are homologous in sequences but disparate in function, have not been studied yet. Here, we identified an amorphous calcium carbonate-binding protein and an acetylcholine-binding protein in the abalone, Haliotis discus hannai , named hdh-ACCBP and hdh-AChBP, respectively. Studies of hdh-ACCBP indicated that it was a counterpart of pf-ACCBP in gastropods that might function similarly in columnar nacre formation and supersaturated extrapallial fluid. Analysis of hdh-AChBP showed that unlike previously identified AChBP, hdh-AChBP was not only expressed in the nervous system but could also be detected in non-nervous system cells, such as the goblet cells of the mantle pallial. Additionally, its expression patterns during embryo and larval development did not accord with ganglion development. These phenomena indicated that AChBP might play more general roles than just in neuronal synapses transmission. Comparison of hdh-ACCBP and hdh-AChBP revealed that they were quite different in their post-translational modification and oligomerization and that they were controlled under different transcriptional regulation systems, consequently obtaining disparate expression profiles. Our results also implied that ACCBP and AChBP might come from a common ancestor through gene duplication and divergence.