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Evidence for aestivation specific proteins inOtala lactea

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Abstract

Changes in [35S]methionine protein labeling patterns were examined by following incorporation into the acid precipitate protein fraction of land snails,Otala lactea (Müller) (Pulmonata, Helicidae). Labeled proteins were analyzed by SDS polyacrylamide gel electrophoresis and isoelectric focusing columns. Snails in four different physiological states were compared: active controls, short term aestivating snails (injected and allowed to enter aestivation), long term aestivating snails (aestivated for 14 days, injected, and maintained in the aestivating state), and snails aroused after aestivation (aestivated, injected, and aroused). Protein associated radioactivity was measured over a 7 day time course post injection. Autoradiographic analysis of SDS-polyacrylamide gels showed increases in the radioactivity of four proteins: 91 kDa (hepatopancreas, day 1 in long term aestivating animals), 50 kDa (hepatopancreas, day 2 in short term aestivating snails), 70 kDa and 30 kDa (foot, day 2 in short term aestivating animals). Hepatopancreas and foot from day 1 long term aestivating and day 2 short term aestivating animals were also analyzed by isoelectric focusing columns. Several pH-specific differences were apparent when controls and aestivating animals were analyzed. In particular a peak of radioactivity was observed at pH 5.05 in 1 d long term aestivating hepatopancreas and at pH 4.30 in 2d short term aestivating animals. Several differences were noted in foot with no specific pattern emerging. SDS-polyacrylamide gel electrophoresis analysis of the hepatopancreas peaks showed the appearance of several bands with increased radioactivity, including the 91 kDa and 50 kDa proteins described above. These results suggest thatO. lactea aestivation specific proteins may be involved in the transition to a depressed metabolic state.

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Abbreviations

dpm:

radioactive disintegrations per minute

PAGE:

polyacrylamide gel electrophoresis

SDS:

sodium dodecyl sulphate

SRP:

stress related protein

References

  1. Pipkin JL, Anson JF, Hinson WG, Burnr ER, Casciano DA: Comparative immunochemical and structural similarities of five stress proteins from various tissue types. Comp Biochem Physiol 89B: 43–50, 1988

    Google Scholar 

  2. Subjeck JR, Shyy T-T: Stress protein systems of mammalian cells. Am J Physiol 250: C1-C17, 1986

    Google Scholar 

  3. Gething M-J, Sambrook J: Protein folding in the cell. Nature 355: 33–45, 1992

    Google Scholar 

  4. Anderson GR, Farkas BK: The major anoxic stress response protein p34 is a distinct lactate dehydrogenase. Biochemistry 27: 2187–2193, 1988

    Google Scholar 

  5. Nickells RW, Browder LW: A role for glyceraldehyde 3-phosphate dehydrogenase in the development of thermotolerance in Xenopus laevis embryos. J Cell Biol: 107: 1901–1909, 1988

    Google Scholar 

  6. Nickells RW, Browder LW, Wang TI: Factors influencing the heat shock response of Xenopus laevis embryos. Can J Biochem Cell Biol 67: 687–695, 1989

    Google Scholar 

  7. Atkinson BG, Pollock M: Effect of heat shock on gene expression in human epidermoid carcinoma cells (strain KB) and in primary cultures of mammalian and avian cells. Can J Biochem 60: 316–327, 1982

    Google Scholar 

  8. Kelley PM, Schlesinger MJ: Antibodies to two major chicken heat shock protein cross-react with similar protein in widely divergent species. Mol Cell Biol 2: 267–274, 1982

    Google Scholar 

  9. Margulir BA, Antropova OY, Kharazova AD: 70 kDa heat shock proteins from mollusc and human cells have common structural and functional domains. Comp Biochem Physiol 94B: 621–623, 1989

    Google Scholar 

  10. Ohtsuka K, Masuda A, Nakai A, Nagata K: A novel 40-kDa protein induced by heat shock and other stresses in mammalian and avian cells. Biochem Biophys Res Comm 166: 642–647, 1990

    Google Scholar 

  11. Whyard S, Wyatt GR, Walker VK: The heat shock response inLocusta migratorai. J Comp Physiol B 156: 813–817, 1986

    Google Scholar 

  12. Fujio N, Hatayama T, Kinoshita H, Yukioka M: Induction of four heat-shock proteins and their mRNAs in rat after whole-body hyperthermia. J Biochem 101: 181–187, 1987

    Google Scholar 

  13. Curle CA, Kapoor M: Expression of heat shock genes of Neurospora crassa: effort of hyperthermia and other stress on mRNA levels. Can J Biochem Cell Biol 66: 81–92, 1988

    Google Scholar 

  14. Storey KB: Suspended animation: the molecular basis of metabolic depression. Can J Zool 66: 124–132, 1988

    Google Scholar 

  15. Machin J: Water relationships. In: V. Fretter, J Peake (eds) The Pulmonates, Academic Press, New York, 1975, pp 105–163

    Google Scholar 

  16. Barnhart MC: Respiratory gas tensions and gas exchange in active and dormant land snails:Octa lactea. Physiol Zool 59: 733–745, 1986

    Google Scholar 

  17. Barnhart MC, McMahon BR: Discontinuous CO2 release and metabolic depression in dormant land snails. J Exp Biol 128: 123–138, 1987

    Google Scholar 

  18. Rees BB, Hand SC: Heat dissipation gas exchange and acid-base status in the land snail Oreohelix during short-term estivation. J Exp Biol 152: 77–92, 1990

    Google Scholar 

  19. Storey KB, Storey JM: Metabolic rate depression and biochemical adaptation in anaerobiosir hibernation and estivation. Quart Rev Biol 65: 145–174, 1990

    Google Scholar 

  20. Whitwam RE, Storey KB: Pyruvate kinase from the land snailOtala lactea: regulation by reversible phosphorylation during estivation and anoxia. J Exp Biol 154: 321–337, 1990

    Google Scholar 

  21. Whitwam RE, Storey KB: Regulation of phosphofructokinase during estivation and anoxia in the land snailOtala lactea. Physiol Zool 64: 595–610, 1991

    Google Scholar 

  22. Brooks SPJ, Storey KB: Properties of pyruvate dehydrogenase from the land snailOtala lactea: control of enzyme activity during estivation. Physiol Zool 65: 620–633, 1992

    Google Scholar 

  23. Brooks SPJ, Storey KB: Glycolytic enzyme binding and metabolic control in estivation and anoxia in the land snailOtala lactea. J Exp Biol 151: 193–204, 1990

    Google Scholar 

  24. Brooks SPJ, Storey KB: Patterns of protein synthesis and phosphorylation during anoxia in the land snailOtala lactea. Can J Zool 72: 856–862, 1994

    Google Scholar 

  25. Rutledge PS, Easton DP, Spotila JR: Heat shock proteins from the lungless salamandersEurycea bislineata andDesmognathus ochrophaeus. Comp Biochem Physiol 88B: 13–18, 1987

    Google Scholar 

  26. Bienz M: Developmental control of the heat shock response in Xenopus. Proc Nat Acad Sci (USA) 81: 3138–3142, 1984

    Google Scholar 

  27. Kothary RK, Candido EPM: Induction of a novel set of polypeptides by heat shock or sodium arsenite in cultured cells of rainbow troutSalmo gaidnerii. Can J Biochem 60: 347–355, 1982

    Google Scholar 

  28. Hallberg RL, Kraur KW, Hallberg EM: Induction of acquired thermotolerance in Tetrahymena thermophila: effects of proteins labeling inhibitors. Mol Cell Biol 5: 2061–2069, 1985

    Google Scholar 

  29. Velazquez JM, Lindquist S: Hsp 70: nuclear concentration during environmental stress and cytoplasmic storage during recovery. Cell 36: 655–662, 1984

    Google Scholar 

  30. Pelham HRB: Speculations on the functions of the major heat shock and glucose-regulated proteins. Cell 46: 959–961, 1986

    Google Scholar 

  31. Lai BT, Chin NW, Stanek AE, Keh W, Lankr KW: Quantitation and intracellular localization of the 85K heat shock protein by using monoclonal and polyclonal antibodies. Mol Cell Biol 4: 2802–2810, 1984

    Google Scholar 

  32. Lipshich LA, Cutt JR, Brugge JS: Association of the transforming proteins ofRour Fujinami Y73 avian sarcoma viruses with the same two cellular proteins. Mol Cell Biol 2: 875–880, 1982

    Google Scholar 

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Author notes

  1. S. P. J. Brooks

    Present address: Nutrition Research Division, Food Directorate Health Protection Branch, Health Canada, Banting Research Centre, Tunney's Pasture, K1A 0LZ, Ottawa, Ontario, Canada

Authors and Affiliations

  1. Department of Biology and Institute of Biochemistry, Carleton University, K1S 5B6, Ottawa, Ontario, Canada

    S. P. J. Brooks & K. B. Storey

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  1. S. P. J. Brooks
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  2. K. B. Storey
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Brooks, S.P.J., Storey, K.B. Evidence for aestivation specific proteins inOtala lactea . Mol Cell Biochem 143, 15–20 (1995). https://doi.org/10.1007/BF00925922

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  • DOI: https://doi.org/10.1007/BF00925922

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