Summary

We examined the seasonal variation in environmentally induced protein damage in natural populations of the intertidal mussel Mytilus trossulus. In order to compare the state of protein pools during seasonal variations in environmental temperature, we used solid-phase immunochemical analysis to quantify ubiquitin conjugate concentrations and relative levels of the stress protein hsp70. The two biochemical indices were selected for their cellular roles in irreversible and reversible protein denaturation, respectively. Proteins that are ubiquitinated are irreversibly damaged and are degraded by intracellular proteases; stress proteins act as molecular chaperones to re-fold thermally denatured proteins and, thus, indicate degrees of reversible protein damage. Comparisons involved mussels collected in February and August from two study sites: an intertidal site which subjected animals to a wide range of body temperatures (from approximately 10 to 35 C in summer), and a subtidal site where animals remained submerged throughout the tidal cycle. Our results show that quantities of ubiquitin conjugates and hsp70 were greater in gill tissue from summer-collected mussels than in gills of winter-collected specimens. Ubiquitin conjugate and hsp70 levels were also greater in mussels collected from an intertidal location than in mussels from a submerged population. Our results show that the high summer temperatures normally experienced in the field are sufficient to cause increased denaturation of cellular proteins. Despite increases in the concentrations of heat shock proteins in summer-acclimatized mussels, elevated levels of irreversibly denatured, i.e. ubiquitinated, proteins were still observed, which indicates that the heat shock response may not be able to rescue all heat-damaged proteins. The energy costs associated with replacing heat-damaged proteins and with maintaining the concentrations and activities of heat shock proteins may contribute substantially to cellular energy demands. These increased energy demands may have an impact on the ecological energetic relationships of species, e.g. in the allocations of energy for growth and reproduction, and, as a consequence, may contribute to determining their distribution limits.