Abstract
A set of normal biochemical indices of the hemolymph plasma in the gastropod pulmonate mollusc Achatina fulica was described in this study. A comparative analysis of the whole plasma and its subfractions enriched with and depleted of the oxygen-carrying protein hemocyanin was carried out using spectrophotometry and spectrofluorimetry. Individual features of the absorption spectra were analyzed using fourth derivatives. An optimal method to estimate the protein concentration was identified. The buffer capacity, equivalence points and pK values of the dominant buffer groups were calculated to characterize the acid-base properties of hemolymph plasma and its subfractions. A major role in maintaining the hemolymph buffer capacity was shown to belong to the bicarbonate buffering system. The results of this study are compared with the data for Helix pomatia available in literature. In the future, the tested indices will be used to devise ecotoxicological criteria for environmental assessment.
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Eropkin, M.Yu. and Kuleva, N.V., Biokhimicheskie metody v toksikologii i ekotoksikologii (Biochemical Methods in Toxicology and Ecotoxicology), St. Petersburg, 2008.
Vladimirov, V.G., Lyzlova, S.N., Lyanguzov, A.Yu., Antipenko, A.E., Vasil’eva, I.N., Voinova, N.E., Dizhe, G.P., Komkova, A.I., Kokryakov, V.N., Krasovskaya, I.E., Leonova, L.E., Lyzlova, L.V., Perestenko, O.V., Petrova, T.A., Rezyapkin, V.I., Slepenkov, S.V., Chesnokova, L.S., Sharova, L.A., and Yankovskii, O.Yu., Fermenty i Nukleinovye Kisloty (Enzymes and Nucleic Acids), St. Petersburg, 1997.
Lowry, O.H., Rosebrough, N.J., Farr, A.L., and Randall, R.J., Protein measurement with the Folin phenol reagent, J. Biol. Chem., 1951, vol. 193, pp. 265–275.
Chen, J.T., Shen, S.T., Chung, C.S., Chang, H., Wang, S.M., and Li, N.C., Achatina fulica hemocyanin and its interactions with imidazole, potassium cyanide, and fluoride as studied by spectrophotometry and nuclear magnetic resonance and resonance raman spectrometry, Biochem., 1979, vol. 18, pp. 3097–3101.
Talsky, G., Derivative Spectrophotometry, Weinheim, 1994.
Alyakrinskaya, I.O., Gemoglobiny i Gemotsianiny Bespozvonochnykh. Biokhimicheskie Adaptatsii k Usloviyam Sredy (Hemoglobins and Hemocyanins of Invertebrates. Biochemical Adaptations to Environmental Conditions), Moscow, 1979.
Decker, H., Hellmann, N., Jaenicke, E., Lieb, B., Meissner, U., and Markl, J., Minireview: recent progress in hemocyanin research, Integr. Comp. Biol., 2007, vol. 47, pp. 631–644.
Markl, J., Evolution of molluscan hemocyanin structures, Biochim. Biophys. Acta, 2013, vol. 1834, pp. 1840–1852.
van Holde, K.E. and Miller, K.I., Haemocyanins, Quart. Rev. Biophys., 1982, vol. 15, pp. 1–129.
Boysen, R.I. and Hearn, M.T.W., High-Performance Liquid Chromatography of Peptides and Proteins, Amino Acids, Peptides and Proteins in Organic Chemistry, Hughes, A.B., Ed., Weinheim, 2012, vol. 5, pp. 167–207.
Drexel, R., Siegmund, S., Schneider, H.-J., Linzen, B., Gielens, C., Preaux, G., Lontie, R., Kellermann, J., and Lottspeich, F., Complete amino-acid sequence of a functional unit from a molluscan hemocyanin (Helix pomatia), Biol. Chem., 1987, vol. 368. pp. 617–635.
Burton, R.F., Buffers in the blood of the snail, Helix pomatia L., Comp. Biochem. Physiol., 1969, vol. 29, pp. 919–930.
Brockelman, C.R., Effects of parasitism and stress on hemolymph protein of the African giant snail, Achatina fulica, Z. Parasitenkd., 1978, vol. 57, pp. 137–144.
Lakowicz, J.R., Principles of Fluorescence Spectroscopy, Baltimore, 2006.
Kovacheva, N.P. and Aleksandrova, E.N., Gematologicheskie pokazateli kak indikatory fiziologisheskogo sostoyaniya dekapod: kamchatskogo kraba Paralithodes camtschaticus i rechnykh rakov rodov Astacus i Pontastacus (Hematological Parameters as Indicators of Physiological State of Decapods: Kamchatka Crab Paralithodes camtschaticus and River Crawfish of Astacus and Pontastacus Genus), Moscow, 2010.
Roach, D.K., Analysis of the haemolymph of Arion ater L. (Gastropoda: Pulmonata), J. Exp. Biol., 1963, vol. 40, pp. 613–623.
Matsumoto, M., Morimasa, T., Takeuchi, H., Mori, A., Kosaka, M., Kobayashi, J., and Morii, F., Amounts of inorganic ions and free amino acids in hemolymph and ganglion of the African giant snail, Achatina fulica Ferussac, Comp. Biochem. Physiol. A: Comp. Physiol., 1974, vol. 48, pp. 465–470.
Zhuravlev, V., Bugaj, V., Kodirov, S., Safonova, T., and Staruschenko, A., Giant multimodal heart motoneurons of Achatina fulica: a new cardioregulatory input in pulmonates, Comp. Biochem. Physiol. A: Mol. Integr. Physiol., 2001, vol. 130, pp. 183–196.
Shabelnikov, S.V., Bystrova, O.A., Ivanov, V.A., Margulis, B.A., and Martynova, M., Atrial granular cells of the snail Achatina fulica release proteins into hemolymph after stimulation of the heart nerve, J. Exp. Biol., 2009, vol. 212, pp. 3211–3220.
Komarov, F.I., Korovkin, B.F., and Men’shikov, V.V., Biokhimicheskie Issledovaniya v Klinike (Biochemical Investigations in Medical Practice), Moscow, 2001.
Martin, A.W., Stewart, D.M., and Harrison, F.M., Urine formation in a pulmonate land snail, Achatina fulica, J. Exp. Biol., 1965, vol. 43, pp. 99–123.
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Original Russian Text © T.A. Petrova, A.Yu. Lianguzov, N.M. Malygina, 2016, published in Zhurnal Evolyutsionnoi Biokhimii i Fiziologii, 2016, Vol. 52, No. 1, pp. 34—41.
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Petrova, T.A., Lianguzov, A.Y. & Malygina, N.M. Spectral and acid-base properties of hemolymph plasma and its fractions in the gastropod pulmonate mollusc Achatina fulica . J Evol Biochem Phys 52, 37–45 (2016). https://doi.org/10.1134/S002209301601004X
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DOI: https://doi.org/10.1134/S002209301601004X