Structure elucidation of the metal-binding sites in metallothionein by /sup 113/Cd NMR
/sup 113/Cd NMR has been used to determine the structures of the multiple metal-binding sites in the two major isoproteins of metallothionein from mammalian livers (rabbits, calf, and human) and from Scylla serrata hepatopancreas. The native protein isolated from the livers of rabbits that had been subjected to repeated injections of /sup 113/CdCl/sub 2/ contains an appreciable amount of Zn/sup 2 +/ in addition to /sup 113/Cd/sup 2 +/, ranging from 2 to 3 g-atoms of a total metal content of 7 g-atoms/mol of protein. The native Zn/sup 2 +/ can be replaced in vitro with /sup 113/Cd/sup 2 +/ to give a /sup 113/Cd NMR spectrum consisting of eight distinct multiplets in the chemical shift range of 604-670 ppm. The multiplet structure is due to /sup 113/Cd- /sup 113/Cd scalar coupling arising from two-bond interactions between /sup 113/Cd/sup 2 +/ ions linked to one another by bridging cysteine thiolate ligands. Analysis of the /sup 113/Cd spectra by selective homonuclear /sup 113/Cd decoupling techniques showed that both isoproteins of rabbit liver metallothionein contain two separate metal clusters, one containing four Cd/sup 2 +/ ions (cluster A) and the other containing three (cluster A) and the other containing three (cluster B).
- Research Organization:
- Yale Univ., New Haven, CT
- OSTI ID:
- 5606090
- Journal Information:
- Fed. Proc.; (United States), Vol. 41:13
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
METALLOTHIONEIN
RECEPTORS
STRUCTURAL CHEMICAL ANALYSIS
ZINC
CADMIUM 113
CALVES
CATIONS
CHEMICAL BONDS
COPPER
CRUSTACEANS
CYSTEINE
IN VITRO
LABELLED COMPOUNDS
LIVER
MAN
NMR SPECTRA
PANCREAS
RABBITS
TRACER TECHNIQUES
AMINO ACIDS
ANIMALS
AQUATIC ORGANISMS
ARTHROPODS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BODY
CADMIUM ISOTOPES
CARBOXYLIC ACIDS
CATTLE
CHARGED PARTICLES
DIGESTIVE SYSTEM
DOMESTIC ANIMALS
ELEMENTS
ENDOCRINE GLANDS
EVEN-ODD NUCLEI
GLANDS
INTERMEDIATE MASS NUCLEI
INVERTEBRATES
IONS
ISOMERIC TRANSITION ISOTOPES
ISOTOPE APPLICATIONS
ISOTOPES
MAMMALS
METALLOPROTEINS
METALS
NUCLEI
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
ORGANS
PRIMATES
PROTEINS
RADIOISOTOPES
RUMINANTS
SPECTRA
STABLE ISOTOPES
THIOLS
TRANSITION ELEMENTS
VERTEBRATES
YEARS LIVING RADIOISOTOPES
550201* - Biochemistry- Tracer Techniques
560301 - Chemicals Metabolism & Toxicology- Cells- (-1987)