Cold-stable eye lens crystallins of the Antarctic nototheniid toothfish Dissostichus mawsoni Norman.
The eye lenses of the Antarctic nototheniid fishes that inhabit the perennially freezing Antarctic seawater are transparent at -2 degrees C, whereas the cold-sensitive mammalian and tropical fish lenses display cold-induced cataract at 20 degrees C and 7 degrees C, respectively. No cold-cataract occurs in the giant Antarctic toothfish Dissostichus mawsoni lens when cooled to temperatures as low as -12 degrees C, indicating highly cold-stable lens proteins. To investigate this cold stability, we characterised the lens crystallin proteins of the Antarctic toothfish, in parallel with those of the sub-tropical bigeye tuna Thunnus obesus and the endothermic cow Bos taurus, representing three disparate thermal climes (-2 degrees C, 18 degrees C and 37 degrees C, respectively). Sizing chromatography resolved their lens crystallins into three groups, alpha/betaH, beta and gamma, with gamma crystallins being the most abundant (>40%) lens proteins in fish, in contrast to the cow lens where they comprise only 19%. The upper thermal stability of these crystallin components correlated with the body temperature of the species. In vitro chaperone assays showed that fish alpha crystallin can protect same-species gamma crystallins from heat denaturation, as well as lysozyme from DTT-induced unfolding, and therefore are small Heat Shock Proteins (sHSP) like their mammalian counterparts. Dynamic light scattering measured an increase in size of alphagamma crystallin mixtures upon heating, which supports formation of the alphagamma complex as an integral part of the chaperone process. Surprisingly, in cross-species chaperone assays, tuna alpha crystallins only partly protected toothfish gamma crystallins, while cow alpha crystallins completely failed to protect, indicating partial and no alphagamma interaction, respectively. Toothfish gamma was likely to be the component that failed to interact, as the supernatant from a cow alpha plus toothfish gamma incubation could chaperone cow gamma crystallins in a subsequent heat incubation, indicating the presence of uncomplexed cow alpha. This suggests that the inability of toothfish gamma crystallins to fully complex with tuna alpha, and not at all with the cow alpha crystallins, may have its basis in adaptive changes in the protein that relate to the extreme cold-stability of the toothfish lens.
Kiss
Andor J
AJ
Department of Animal Biology, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801, USA.
Mirarefi
Amir Y
AY
Ramakrishnan
Subramanian
S
Zukoski
Charles F
CF
Devries
Arthur L
AL
Cheng
Chi-Hing C
CH
D000064:Acclimatization / Q000502:physiology*; D000818:Animals; D000864:Antarctic Regions; D002417:Cattle / Q000502:physiology; D002850:Chromatography, Gel; D003080:Cold Temperature; D004591:Electrophoresis, Polyacrylamide Gel; D006360:Heat-Shock Proteins / Q000737:chemistry / Q000502:physiology; D015151:Immunoblotting; D007908:Lens, Crystalline / Q000737:chemistry* / Q000502:physiology; D018832:Molecular Chaperones / Q000378:metabolism; D010473:Perciformes / Q000502:physiology*; D013045:Species Specificity; D038201:alpha-Crystallins / Q000737:chemistry* / Q000502:physiology
D006360:Heat-Shock Proteins; D018832:Molecular Chaperones; D038201:alpha-Crystallins