In-depth proteomic analyses of Haliotis laevigata (greenlip abalone) nacre and prismatic organic shell matrix

Abstract

BackgroundThe shells of various Haliotis species have served as models of invertebrate biomineralization and physical shell properties for more than 20 years. A focus of this research has been the nacreous inner layer of the shell with its conspicuous arrangement of aragonite platelets, resembling in cross-section a brick-and-mortar wall. In comparison, the outer, less stable, calcitic prismatic layer has received much less attention. One of the first molluscan shell proteins to be characterized at the molecular level was Lustrin A, a component of the nacreous organic matrix of Haliotis rufescens. This was soon followed by the C-type lectin perlucin and the growth factor-binding perlustrin, both isolated from H. laevigata nacre, and the crystal growth-modulating AP7 and AP24, isolated from H. rufescens nacre. Mass spectrometry-based proteomics was subsequently applied to to Haliotis biomineralization research with the analysis of the H. asinina shell matrix and yielded 14 different shell-associated proteins. That study was the most comprehensive for a Haliotis species to date.MethodsThe shell proteomes of nacre and prismatic layer of the marine gastropod Haliotis laevigata were analyzed combining mass spectrometry-based proteomics and next generation sequencing.ResultsWe identified 297 proteins from the nacreous shell layer and 350 proteins from the prismatic shell layer from the green lip abalone H. laevigata. Considering the overlap between the two sets we identified a total of 448 proteins. Fifty-one nacre proteins and 43 prismatic layer proteins were defined as major proteins based on their abundance at more than 0.2% of the total. The remaining proteins occurred at low abundance and may not play any significant role in shell fabrication. The overlap of major proteins between the two shell layers was 17, amounting to a total of 77 major proteins.ConclusionsThe H. laevigata shell proteome shares moderate sequence similarity at the protein level with other gastropod, bivalve and more distantly related invertebrate biomineralising proteomes. Features conserved in H. laevigata and other molluscan shell proteomes include short repetitive sequences of low complexity predicted to lack intrinsic three-dimensional structure, and domains such as tyrosinase, chitin-binding, and carbonic anhydrase. This catalogue of H. laevigata shell proteins represents the most comprehensive for a haliotid and should support future efforts to elucidate the molecular mechanisms of shell assembly.Electronic supplementary materialThe online version of this article (10.1186/s12953-018-0139-3) contains supplementary material, which is available to authorized users.

Mapping the Tooth Enamel Proteome and Amelogenin Phosphorylation Onto Mineralizing Porcine Tooth Crowns

“…The spatial distributions of AMELX, AMBN, ENAM, MMP20, and KLK4 (Figures 1D, 4) agree very well with published data on abundance patterns during specific stages of enamel formation (see Moradian-Oldak, 2012; Bartlett, 2013; Lacruz et al, 2017 and references therein). Our findings are consistent with the number and identifications of proteins of recently published proteomics analyses of pooled human and rodent enamel (Chen et al, 1995; Hubbard and Kon, 2002; Jagr et al, 2012, 2014; Eckhardt et al, 2014; Charone et al, 2016; Pandya et al, 2017; De Lima Leite et al, 2018; Mann et al, 2018; Zhang et al, 2018). For instance, we mapped 19 of the 24 proteins recently reported in enamel onto enamel formation stages (Pandya et al, 2017), and localized close homologs for the remaining five proteins (Supplementary Table 1).…”

Molecular modularity and asymmetry of the molluscan mantle revealed by a gene expression atlas

“…BLASTp-based comparisons of the L. stagnalis shell proteome were performed against a variety of calcifying proteomes reported in a wide phylogenetic range of metazoans as described in [ 14 ]. These included 42 proteins from the oyster Pinctada maxima reported in [ 47 ]; 78 proteins from the oyster Pinctada margaritifera reported in [ 47 ]; 94 proteins from the abalone Haliotis asinina reported in [ 17 ] and [ 16 ]; 80 proteins from the abalone H. laevigata reported in [ 48 ]; 63 proteins from the limpet Lottia gigantea reported in [ 49 ]; 53 proteins from the oyster Crassostrea gigas reported in [ 50 ]; 71 proteins from the mussel Mya truncata reported in [ 51 ]; 59 proteins from the grove snail Cepaea nemoralis reported in [ 14 ]; 44 proteins from the oyster Pinctada fucata reported in [ 52 ]; 53 proteins from the mussel Mytilus coruscus reported in [ 53 ]; 66 proteins from the brachiopod Magellania venosa reported in [ 54 ]; 139 proteins from the sea urchin Strongylocentrotus purpuratus reported in [ 55 ]; and 37 proteins from the coral Acropora millepora reported in [ 56 ].…”

A Novel Tyrosinase Gene Plays a Potential Role in Modification the Shell Organic Matrix of the Triangle Mussel Hyriopsis cumingii

“…Tyrosinases catalyze the oxidation of both monophenols and o-diphenols into reactive o-quinones (Garcia-Borron and Solano, 2002). Data from shell proteomics showed that tyrosinases were incorporated into the shell matrices in the prismatic layer (Nagai et al, 2007;Liao et al, 2015;Mann et al, 2018), nacre layer (Mann et al, 2018), and even the whole shell (Zhang et al, 2012;Du et al, 2017) of molluscan species. Recent research showed that tyrosinases participated in periostracum sclerotization and the cross-linking and maturation of shell matrix proteins by catalyzing the quinoprotein oxidation (Zhang et al, 2012;Aguilera et al, 2014;Du et al, 2017).…”

“…Recent research has shown that tyrosinases are strongly expressed in the pallial and edge of the mantle, which are the main biomineralization tissues of Mollusca (Zhang et al, 2006a;Nagai et al, 2007;Yu et al, 2014;Huening et al, 2016;Yang et al, 2017), and tyrosinase proteins have also been detected in the proteome of the shell prismatic layer and/or nacreous layer (Marie et al, 2012;Liao et al, 2015;Mann et al, 2018). These results implied the crucial functions of tyrosinases in periostracum sclerotization and the cross-linking and maturation of shell matrix proteins (Zhang et al, 2012;Aguilera et al, 2014;Huening et al, 2016;Du et al, 2017).…”