Shotgun analysis of the marine mussel Mytilus edulis hemolymph proteome and mapping the innate immunity elements

Abstract

Shotgun analysis of the marine mussel Mytilus edulis hemolymph proteome and mapping the innate immunity elements. Campos A, Apraiz I, da Fonseca RR, Cristobal S. Proteomics 2015, 15, 4021-4029 DOI 10.1002 1 AbstractThe marine mussel innate immunity provides protection to pathogen invasion and inflammation. In this regard the mussel haemolymph takes a main role in the animal innate response. Despite the importance of this body fluid in determining the physiological condition of the animal, little is known about the molecular mechanisms underlying the cellular and humoral responses. In this work we have applied a mass spectrometry (nanoLC-MS/MS) strategy integrating genomic and transcriptomic data with the aim to: i) identify the main protein functional groups that characterize haemolymph and ii) to map the elements of innate immunity in the marine mussel Mytilus edulis haemolymph proteome.After sample analysis and first protein identification based on MS/MS data comparison, proteins with unknown functions were annotated with blast using public database (nrNCBI) information. Overall 595 haemolymph proteins were identified with high confidence and annotated. These proteins encompass primary cellular metabolic processes: energy production and metabolism of biomolecules, as well as processes related to oxidative stress defence, xenobiotic detoxification, drug metabolism, and immune response. A group of proteins was identified with putative immune effector, receptor and signalling functions in M. edulis. Data are available via ProteomeXchange with identifier PXD001951. Significance of the Study:Proteomic investigations have been hampered in Mytilus sp. by the scarce genomic data available.Here a high-throughput methodology was successfully employed to describe the overlooked haemolymph proteome from the marine mussel M. edulis, revealing a conjunct of molecular functions contributing to the immune defence. The information available contributes to enhance the Shotgun analysis of the marine mussel Mytilus edulis hemolymph proteome and mapping the innate immunity elements. Campos A, Apraiz I, da Fonseca RR, Cristobal S.

Shift in Immune Parameters After Repeated Exposure to Nanoplastics in the Marine Bivalve Mytilus

“…EPp, the most abundant serum protein in M. galloprovincialis, is an acidic, histidine-rich, cation binding glycoprotein; it has a complex and anomalous N-glycan structure and contains a conserved C1q complement domain. Due to its peculiar composition, EPp is involved in multiple functions, from shell formation to immune response (42)(43)(44). This protein has been shown to play a key role in specific recognition of both selected bacterial strains and NP types.…”

Proteomic Analyses of the Unexplored Sea Anemone Bunodactis verrucosa

“…A methodology based on shotgun analysis was employed to investigate the whole-body proteome of B. verrucosa . This methodology has been previously reported as suitable for diverse purposes related to protein identification such as characterization of complex sample, inference of the main enzymatic pathway involved in a tissue, even to reveal venom composition [ 68 , 69 , 70 , 71 ]. Altogether, 688 peptide sequences were identified among the two replicates of the fractions analyzed (SF and IF), which accounted for 412 groups of non-redundant proteins (), retrieved from custom cnidarians databases.…”

“…FASP protein digests (duplicate samples) were analyzed by nano-LC coupled to a hybrid Ion-trap mass spectrometer (LTQ Orbitrap Velos Pro—ETD, Thermo Scientific) as described previously [ 68 ]. Peptides were separated by reverse-phase chromatography (20 mm × 100 µm C18 precolumn followed by a 100 mm × 75 µm C18 column with particle size 5 µm, NanoSeparations, Nieuwkoop, The Netherlands) using a linear ascending gradient of buffer B (ACN + FA, 0.1%, v / v ), being buffer A TFA, 0.1%, v / v in water.…”

Top-Down Proteomics and Farm Animal and Aquatic Sciences

“…Interest at the level of gene and protein function will provide a more comprehensive understanding of a species’ biology and its response/adaptation to the environment. The main areas of research that have adhered to this technology have been aquaculture for improving fish health and nutrition, welfare assessment and stress reduction, diseases and the use of antibiotics and vaccines, aquatic toxicology, and the identification of major environmental threats and the mode of action (MOA) of water contaminants [52,53,54]; developmental biology and reproduction [55,56,57]; and physiology and immune mechanisms in selected wildlife and commercial species [58,59,60,61]. The conventional proteomic techniques such as quantitative 2DE and 2D-DIGE in combination with MS have been the most used to address specific proteomic questions raised in aquatic sciences.…”