Purification and characterization studies of cadmium-binding proteins from the American oyster, Crassostrea virginica.
The previously reported low molecular weight cadmium-binding protein (CdBP) from the American oyster, Crassostrea virginica, has been further purified and characterized by improved technical methods. The internal organ distribution of the protein within the oyster and effects of life cycle/season on CdBP production also have been evaluated. CdBP isolated by extended ion-exchange gradients or double ion-exchange chromatography followed by HPLC analysis possesses an electrophoretic Rf of about 0.7 and contains relatively little Zn, as previously reported. Cysteine, lysine, and glycine are the dominant amino acids. When ion-exchange columns are developed with NaCl gradients, the aromatic residues tryptophan, tyrosine, and phenylalanine are found to be present, but these may be largely removed depending upon whether the protein is denatured and carboxymethylated prior to analysis. The ultraviolet absorption spectrum of CdBP also was variable, with 250/280 nm ratios ranging from 17:1 immediately after ion-exchange chromatography to 2:1 following concentration procedures. Internal organ distribution studies showed that the visceral mass contained most of the Cd present with lesser amounts in the gills and mantle. In contrast with mammals, CdBP accounts for only about 30% of the total cell Cd burden in these tissues. Cu displacement of Cd from the protein is a particular problem during the summer spawning season and appears to stem from altered Cu metabolism during this period. Relative oyster dormancy during the winter also reduces CdBP production in response to Cd, and the protein is obtained most readily during the fall and spring.(ABSTRACT TRUNCATED AT 250 WORDS)
Fowler
B A
BA
Engel
D W
DW
Brouwer
M
M
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002104:Cadmium / Q000378:metabolism*; D002850:Chromatography, Gel; D003300:Copper / Q000378:metabolism; D003545:Cysteine / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D008668:Metallothionein / Q000302:isolation & purification* / Q000378:metabolism; D010125:Ostreidae / Q000032:analysis* / Q000378:metabolism; D012621:Seasons; D013056:Spectrophotometry, Ultraviolet; D015032:Zinc / Q000378:metabolism
D000596:Amino Acids; C004203:cadmium-binding protein; D002104:Cadmium; D003300:Copper; D008668:Metallothionein; D015032:Zinc; D003545:Cysteine