Cell proteome variability of protistan mollusc parasite Perkinsus olseni among regions of the Spanish coast

Abstract

We evaluated the proteome variability of in vitro-cultured Perkinsus olseni cells deriving from 4 regions of the Spanish coast: the rías of Arousa and Pontevedra (Galicia, N W Spain), Carreras River in Huelva (Andalusia, SW Spain) and Delta de l'Ebre (Catalonia, N E Spain). P. olseni in vitro clonal cultures were produced starting from parasite isolates from 4 individual clams from each region. Those clonal cultures were used to extract cell proteins, which were separated by 2-dimensional (2D) electrophoresis. Qualitative comparison of P. olseni protein expression profiles among regions was performed with PD Quest software. Around 700 protein spots from parasites derived from each region were considered, from which 141 spots were shared by all the regions. Various spots were found to be exclusive to each region. Higher similarity was found among the proteomes of P. olseni from the Atlantic regions than between those from the Mediterranean and the Atlantic. A total of 54 spots were excised from the gels and sequenced. Nineteen proteins were annotated after searching in databases, 13 being shared by all the regions and 6 exclusive to 1 region. Most of the identified proteins were involved in glycolysis, oxidation/ reduction, metabolism and response to stress. No direct evidence of P. olseni variability associated with virulence was found within the protein set analysed, although the differences in metabolic adaptation and stress response could be connected to pathogenicity. KEY WORDS: Ruditapes decussatus · Ruditapes philippinarum · Mollusc · 2D electrophoresis · In vitro cultureResale or republication not permitted without written consent of the publisher

New insights into the Manila clam – Perkinsus olseni interaction based on gene expression analysis of clam hemocytes and parasite trophozoites through in vitro challenges

“…olseni trophozoite, such as AHPC, a member of family peroxidases involved in controlling endogenous and exogenous peroxides in response to stress (Hofmann et al, 2002), or GST, peroxiredoxin 5 and membrane selenoprotein, previously related to antioxidant activity in Perkinsus spp. (Araujo et al, 2013, Fernández-Boo et al, 2014, 2015a, 2015b. On the other hand, the downregulation in the trophozoite of prostatic acid phosphatase precursor might hinder the ability of P.…”

“…Clam perkinsosis has also been studied from the parasite point perspective focusing on different biological processes, including its proliferation (Casas et al, 2002b;Elandalloussi et al, 2003Elandalloussi et al, , 2005aAraujo et al, 2013), metabolic pathways (Elandalloussi et al, 2005b), pathogenicity (Shimokawa et al, 2010;Waki et al, 2012;Waki and Yoshinaga, 2013), population genetic structure (Pardo et al, 2011;Vilas et al, 2011), and gene and protein expression profiles (Ascenso et al, 2007;Leite et al, 2008;Ascenso, 2011;Fernández-Boo et al, 2014, 2015a, 2015b.…”

Shotgun Label-Free Proteomic Analyses of the Oyster Parasite Perkinsusmarinus

“…Additional proteins that are important for the pathogenesis of P. marinus and a variety of bacterial and parasitic organisms were detected in at least two replicates, including: aldose reductase [20], betaine aldehyde dehydrogenase [21], helicases [15,22], malate dehydrogenase [6], and aspartyl aminopeptidase [23].…”

“…In contrast, few studies available on comparisons of proteomic profiles of different Perkinsus species and on proteome variability of P. olseni resulted in low throughputs: 28 and 19 annotated proteins, respectively [5,6]. More proteomic baseline data is required for a better understanding of P. marinus biology, including virulence mechanisms.…”