Optimization of skeletal protein preparation for LC-MS/MS sequencing yields additional coral skeletal proteins in Stylophora pistillata

ABSTRACT: Stony corals generate their calcium carbonate exoskeleton in a highly controlled biomineralization process mediated by a variety of macromolecules including proteins. Fully identifying and classifying these proteins is crucial to understanding their role in exoskeleton formation, yet no optimal method to extract and isolate and characterize coral skeletal proteins has been established and their complete composition remains obscure. Here, we tested four skeletal protein extraction protocols using acetone precipitation and ultrafiltration dialysis filters to present a comprehensive scleractinian coral skeletal proteome. We identified a total of 60 proteins in the coral skeleton, 44 of which were not present in previously published stony coral skeletal proteomes. Extracted protein treatment protocols carried out in this study revealed that there is no “one optimal method” and each protocol revealed a unique set of method-exclusive proteins. To better understand the general mechanism of skeletal protein transportation, we further examined the proteins’ gene ontology, transmembrane domains, and signal peptides. We found that transmembrane domain proteins and signal peptide secretion pathways, by themselves, could not explain the transportation of proteins to the skeleton . We therefore propose that proteins are transported to the skeletal via vesicles and possibly non-traditional secretion pathways.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Stylophora Pistillata Anthozoa

SUBMITTER: David Morgenstern  

LAB HEAD: Tali Mass

PROVIDER: PXD017891 | Pride | 2020-06-16

REPOSITORIES: Pride