Purification and characterization of a cadmium-induced metallothionein from the shore crab Carcinus maenas (L.).
Two metallothionein variants were purified from the midgut gland of crabs (Carcinus maenas) exposed to a high cadmium concentration (2 p.p.m.). One of the variants was purified from crabs exposed to a low cadmium concentration (0.5 p.p.m.). The purification method involved acetone precipitation, gel filtration and reversed-phase h.p.l.c. The complete amino acid sequences of both variants have been elucidated by m.s. and automated sequence analysis on S-methylated proteins or fragments produced by cleavage of the S-methylated proteins with Staphylococcus aureus proteinase. The two variants from crabs exposed to the high cadmium concentration differed only by a single residue of methionine at the N-terminus. The single variant isolated from crabs exposed to the low cadmium concentration was the one without the N-terminal methionine, indicating that high cadmium concentrations either inhibit the processing enzymes and/or that the processing enzymes cannot keep pace with the increased metallothionein synthesis when cadmium availability is high. Cadmium-induced metallothionein from C. maenas shows a high degree of structural similarity to metallothioneins from the decapod crustaceans Scylla serrata and Homarus americanus.
Pedersen
K L
KL
Institute of Biology, University of Odense, Denmark.
Pedersen
S N
SN
Højrup
P
P
Andersen
J S
JS
Roepstorff
P
P
Knudsen
J
J
Depledge
M H
MH
D000595:Amino Acid Sequence; D000818:Animals; D003386:Brachyura / Q000378:metabolism*; D002104:Cadmium / Q000494:pharmacology*; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D008297:Male; D013058:Mass Spectrometry / Q000379:methods; D008668:Metallothionein / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification*; D008969:Molecular Sequence Data
D002104:Cadmium; D008668:Metallothionein