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Copper isotope composition of hemocyanin

Abstract :
Background: Copper is a metal that plays a central role in biology, for example, as co-factor in various redox enzymes. Its stable isotopic composition is being used as tracer of its transport in living organisms and as a biomarker for diseases affecting its homeostasis. While the application of copper stable isotopes to biological studies is a growing field, there are presently no biological standards that are systematically analyzed in the different laboratories, as it is the case for geological samples (e.g., by using widely available basalt samples). It is therefore paramount for the community to establish such standard. Copper also binds oxygen in the respiratory protein, hemocyanin, in the hemolymph of mollusks and arthropods and is thus critical to respiration for these species.

Methods: Here, the Cu isotope composition of hemocyanin of different modern species of mollusks and arthropods (Megathura crenulate Keyhole limpet, Limulus polyphemus Horseshoe crab and Concholepas concholepas Chilean abalone), as well as theoretical constraints on the origin of these isotopic fractionations through ab initio calculations are reported.

Results: The isotopic fractionation factors for Cu(I) and Cu(II), both in hemocyanin and in seawater, predict an enrichment in the lighter isotope of Cu in the hemocyanin by over 1 permil compared to seawater. The hemocyanin of Chilean abalone and Horseshoe crab have Cu isotope compositions (δ65Cu = +0.63 ± 0.04‰ and +0.61 ± 0.04‰, respectively, with δ65Cu the permil deviation of the 65Cu/63Cu ratio from the NIST SRM 976 standard), similar to that of the octopus reported in literature (+0.62‰), that are undistinguishable from seawater, suggesting quantitative Cu absorption for these organisms. Conversely, the Keyhole limpet is enriched in the lighter isotope of Cu, which is in line with the ab initio calculation and therefore Cu isotopic fractionation during incorporation of Cu into the hemocyanin.

Conclusions: Because these hemocyanin standard samples are widely available, they could serve in the future as inter-laboratory standards to verify the accuracy of the Cu isotopic measurements on biological matrices.
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Submitted on : Wednesday, May 18, 2022 - 7:02:15 PM
Last modification on : Saturday, June 25, 2022 - 3:43:48 AM

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Marine Paquet, Toshiyuki Fujii, Frédéric Moynier. Copper isotope composition of hemocyanin. Journal of Trace Elements in Medicine and Biology, Elsevier, 2022, 71, pp.126967. ⟨10.1016/j.jtemb.2022.126967⟩. ⟨hal-03671839⟩

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