Cytosolic distribution of copper in the gills of field-collected oysters with different copper bioaccumulation

Abstract

Oysters can hyper-accumulate copper (Cu) without apparent toxicity, but the mechanism of sequestering excessive cytosolic Cu in oysters remains unclear. We here investigated the Cu distribution in the cytosolic proteins (CPs) in the gills of oysters (Crassostrea hongkongensis) through size-exclusion chromatography coupled to inductively coupled plasma mass spectrometry (SEC-ICP-MS). Oysters collected from the southern coast of China contained a gradient of gill Cu concentrations ranging from 132 to 3540 μg g−1 (dry weight), with 7–41 % of Cu distributed in the CPs fraction. The CPs-Cu concentrations were 8.6 times higher in oysters with high Cu concentrations compared to low concentrations. In the CPs, Cu was dispersed with a broad range of molecular weight, suggesting the involvement of various cytosolic proteins in Cu binding. Among the 10 major Cu peaks, peaks 2 (>600 kDa) and peak 8 (18 kDa) contained substantial Cu and showed obvious differences in response to the variation of CPs-Cu levels. Peak 8 contained metallothionein-like proteins that decreased their role in Cu binding as CPs-Cu concentrations increased. LC-MS/MS analysis revealed that peak 2 contained macromolecular protein complexes (MPCs), which played a critical role in binding excess Cu. The comparison with other bivalve species further suggested that sequestering excess CPs-Cu in MPCs was a special strategy employed by oysters in response to high Cu accumulation. This study provides valuable insights into the mechanism of hyper-accumulation and sequestration of Cu in oysters and helps to better understand Cu biomonitoring by oysters.