Copper transport by lobster (Homarus americanus) hepatopancreatic lysosomes

Abstract

Lysosomes are known centers for sequestration of calcium and a variety of heavy metals in many invertebrate tissues, and as a result of this compartmentalization these organelles perform important detoxification roles in the animals involved. The present investigation uses a centrifugation method to isolate and purify hepatopancreatic lysosomes from the American lobster, Homarus americanus. Purified lysosomal preparations were used to characterize membrane transport mechanisms in these organelles for transferring and sequestering cytoplasmic copper following its absorption across the plasma membrane from dietary constituents. The copper-specific fluorescent dye, Phen Green, was employed to quantify transmembrane fluxes of this metal as has been recently used to investigate copper movements across hepatopancreatic mitochondrial and plasma membranes. Results indicated the presence of a vanadate-sensitive, calcium-stimulated, copper ATPase in the membranes of these organelles that displayed high affinity carrier-mediated transport kinetics and may significantly contribute to organismic copper homeostasis. Together with a putative bafilomycin-sensitive V-ATPase in the membrane of the same organelles, importing hydrogen ions into the organellar interior, this copper ATPase may function as part of a physiological mechanism for precipitate formation between metallic cations and anions. These ionic precipitate complexes may then act as a sink for excess metals and thereby reduce the circulating concentrations of these elements.