Isolation and partial characterization of a high molecular weight Cd/Zn-binding protein from the kidney of the scallop Placopecten magellanicus: preliminary studies.
Exposure of the scallop Placopecten magellanicus to 20 ppb Cd2+ in seawater for 7 weeks results in a 7-fold increase in the kidney cytosol content of Cd and 5-fold increase in Zn. Sephadex G-75 column chromatography of the kidney cytosol showed that most of the Cd and Zn were bound to a protein complex with an estimated molecular mass of 45,000 daltons. Further purification of this complex by DEAE A-25 column chromatography disclosed the presence of five peaks with varying degrees of affinity for the ion-exchange resin. One of these peaks (III) was successfully rechromatographed by ion-exchange chromatography and further purified by HPLC using a gel-permeation column. The resultant protein peak which was resistant to disaggregation by 20 mM dithiothreitol gave a preliminary amino acid composition with cysteine, glycine, alanine, and lysine as the major amino acids. The aromatic amino acid phenylalanine was also present. The ultraviolet absorption spectrum gave a 250/280 nm ratio of 2.5:1. Metal analysis of the purified protein showed that it contained Cd, Zn, and Cu in ratios of 1:1:1. Results of these studies indicate that scallop kidney produces a protein complex which appears to share both similarities with mammalian metallothionein with respect to the presence of both Cd and Zn but different with respect to apparent size, amino acid composition, and ultraviolet absorption spectrum.
B A
BA
Fowler
M M
MM
Megginson
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D005075:Biological Evolution; D002104:Cadmium / Q000378:metabolism*; D002352:Carrier Proteins / Q000302:isolation & purification*; D002845:Chromatography; D007668:Kidney / Q000032:analysis; D046911:Macromolecular Substances; D008668:Metallothionein / Q000302:isolation & purification*; D008970:Molecular Weight; D008974:Mollusca / Q000378:metabolism*; D013056:Spectrophotometry, Ultraviolet; D015032:Zinc / Q000302:isolation & purification*
D000596:Amino Acids; D002352:Carrier Proteins; D046911:Macromolecular Substances; C004203:cadmium-binding protein; C017800:zinc-binding protein; D002104:Cadmium; D008668:Metallothionein; D015032:Zinc