Structural characterization and thermal stability of Notothenia coriiceps metallothionein.
Fish and mammalian metallothioneins (MTs) differ in the amino acid residues placed between their conserved cysteines. We have expressed the MT of an Antarctic fish, Notothenia coriiceps, and characterized it by means of multinuclear NMR spectroscopy. Overall, the architecture of the fish MT is very similar to that of mammalian MTs. However, NMR spectroscopy shows that the dynamic behaviour of the two domains is markedly different. With the aid of absorption and CD spectroscopies, we studied the conformational and electronic features of fish and mouse recombinant Cd-MT and the changes produced in these proteins by heating. When the temperature was increased from 20 to 90 degrees C, the Cd-thiolate chromophore absorbance at 254 nm of mouse MT was not modified up to 60 degrees C, whereas the absorbance of fish MT decreased significantly starting from 30 degrees C. The CD spectra also changed quite considerably with temperature, with a gradual decrease of the positive band at 260 nm that was more pronounced for fish than for mouse MT. The differential effect of temperature on fish and mouse MTs may reflect a different stability of metal-thiolate clusters of the two proteins. Such a conclusion is also corroborated by results showing differences in metal mobility between fish and mouse Zn-MT.
D'Auria
S
S
CNR, Institute of Protein Biochemistry and Enzymology, via Marconi 10, I 80125 Naples, Italy.
Carginale
V
V
Scudiero
R
R
Crescenzi
O
O
Di Maro
D
D
Temussi
P A
PA
Parisi
E
E
Capasso
C
C
D000595:Amino Acid Sequence; D000818:Animals; D002104:Cadmium / Q000378:metabolism; D002942:Circular Dichroism; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D008668:Metallothionein / Q000737:chemistry* / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D010473:Perciformes / Q000378:metabolism*; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012118:Resorcinols / Q000378:metabolism; D016415:Sequence Alignment; D013054:Spectrophotometry, Atomic; D013696:Temperature; D015032:Zinc / Q000378:metabolism
D011994:Recombinant Proteins; D012118:Resorcinols; D002104:Cadmium; D008668:Metallothionein; D015032:Zinc; C028398:4-(2-pyridylazo)resorcinol