Fish and molluscan metallothioneins.
Metallothioneins (MTs) are noncatalytic peptides involved in storage of essential ions, detoxification of nonessential metals, and scavenging of oxyradicals. They exhibit an unusual primary sequence and unique 3D arrangement. Whereas vertebrate MTs are characterized by the well-known dumbbell shape, with a beta domain that binds three bivalent metal ions and an alpha domain that binds four ions, molluscan MT structure is still poorly understood. For this reason we compared two MTs from aquatic organisms that differ markedly in primary structure: MT 10 from the invertebrate Mytilus galloprovincialis and MT A from Oncorhyncus mykiss. Both proteins were overexpressed in Escherichia coli as glutathione S-transferase fusion proteins, and the MT moiety was recovered after protease cleavage. The MTs were analyzed by gel electrophoresis and tested for their differential reactivity with alkylating and reducing agents. Although they show an identical cadmium content and a similar metal-binding ability, spectropolarimetric analysis disclosed significant differences in the Cd7-MT secondary conformation. These structural differences reflect the thermal stability and metal transport of the two proteins. When metal transfer from Cd7-MT to 4-(2-pyridylazo)resorcinol was measured, the mussel MT was more reactive than the fish protein. This confirms that the differences in the primary sequence of MT 10 give rise to peculiar secondary conformation, which in turn reflects its reactivity and stability. The functional differences between the two MTs are due to specific structural properties and may be related to the different lifestyles of the two organisms.
Vergani
Laura
L
Department of Biophysical Sciences and Technologies, M. & O. University of Genova, Italy. Laura.Vergani@unige.it
Grattarola
Myriam
M
Borghi
Cristina
C
Dondero
Francesco
F
Viarengo
Aldo
A
D000595:Amino Acid Sequence; D000818:Animals; D008668:Metallothionein / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008670:Metals / Q000737:chemistry; D008969:Molecular Sequence Data; D049878:Mytilus / Q000737:chemistry*; D017686:Oncorhynchus mykiss / Q000378:metabolism*; D010084:Oxidation-Reduction; D011485:Protein Binding; D011487:Protein Conformation; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D013696:Temperature
D008670:Metals; D008668:Metallothionein