Two-dimensional gel electrophoresis of Mytilus galloprovincialis differences in protein expression between intertidal and cultured mussels

Abstract

Two-dimensional gel electrophoresis (2-DE) is a unique method of large-scale protein characterisation, and is a powerful approach in the study of protein expression. In the present work, the experimental conditions for 2-DE of foot proteins from the mussel Mytilus galloprovincialis Lmk were established; the technique was performed with intertidal and cultured mussels using Melanie 3 software for data analysis. This powerful technique enabled the visualisation of a total of 750 protein spots consistently expressed in the foot. The intensity of 92 selected spots was compared between intertidal and cultured mussels, and statistically significant differences were detected in the expression of 45 (48.9%) of the 92 proteins analysed. In 31 of these proteins, intensity was higher in the cultured stock than in the intertidal mussels, while in 14 proteins spot intensity was higher in the latter. Using mass spectrometry (MS) combined with sequence database searching, 6 of the most prominent differentially expressed proteins were analysed. Of these, 1 was identified as being Heat-shock Protein 70, and 2 were shown to be cytoskeleton-associated proteins, myosin and actin. Heat-shock Protein 70, which is known to be involved in cellular transport and chaperoning and associated with stress situations, was more highly expressed in intertidal mussels living in littoral areas than in cultured mussels. These findings are discussed in connection with the molecular changes involved in the adaptation of mussels to different ecological conditions. KEY WORDS: Mytilus galloprovincialis · Two-dimensional gel electrophoresis · Proteome · Foot proteinsResale or republication not permitted without written consent of the publisher

Proteome response to heat stress in the Antarctic clam Laternula elliptica

“…e coe cient of determination (r 2 ) for normalised spot volume for each animal for which technical replicates were run (0.92 to 0.97) was within the range reported in previous studies in bivalves [30,46], indicating that technical variation was consistent with expectations from 2-DE analysis. Figure 1.…”

Proteomics Identification of Azaspiracid Toxin Biomarkers in Blue Mussels, Mytilus edulis

“…For instance, Ló pez et al (22) used proteomics to expand their understanding of the molecular differentiation between the mussels M. edulis and Mytilus galloprovincialis, whereas Apraiz et al (23) identified the proteomic signatures in mussels exposed to marine pollutants.…”

Applications of proteomics in marine ecology

“…In the marine environment, 2DE has been used to screen organisms for the presence of bioactive compounds and to detect and quantify changes in gene expression at the protein level during development, as well as in response to different physiological and environmental conditions (e.g. López et al 2001, 2002, 2005, Olsson et al 2004, Schweder et al 2005, Barneah et al 2006, McDonagh et al 2006. 2DE also has great potential for the study of genetic variability of populations, in that it allows a more representative sample of the genome to be analyzed.…”

“…The use of proteomics to measure changes in the levels of expression at protein level has enabled rapid advances in our understanding of the ecological and environmental adaptations of organisms, as well as of the biogeographical distribution of species (López et al 2001, 2002, Fuentes et al 2002). Proteomics provides a higher level of analysis to aid the understanding of gene function in particular and biology in general (López 2005).…”

“…When working with samples from many marine species, we are limited by the availability of genomics and proteomics resources for those species and, in many instances, even closely related species. For example, López et al (2001) compared differences in protein expression between intertidal and cultured populations of Mytilis galloprovincialis using high resolution 2DE. Over 750 proteins that were consistently expressed in foot tissues were observed in that study.…”