Abstract
A cDNA encoding a second zinc transporter (ZnT-2) was isolated from a rat kidney cDNA expression library by complementation of a zinc-sensitive BHK cell line. The protein predicted from the open reading frame of ZnT-2 cDNA has 359 amino acids and initiates with a CTG codon. It resembles ZnT-1 (a plasma membrane protein that stimulates zinc efflux) in overall topology in that it has six membrane-spanning domains, a histidine-rich intracellular loop and a long C-terminal tail; however, the overall amino acid identity is only 26%. Unlike ZnT-1, which is in the plasma membrane and lowers cellular zinc by stimulating zinc efflux, ZnT-2 is localized on vesicles and allows the zinc-sensitive BHK cells to accumulate zinc to levels that are much higher than non-transformed cells can tolerate. Zinc was visualized within these vesicles with zinquin, a zinc-specific fluorescent probe. The intracellular compartment that accumulates zinc is acidic as revealed by staining with acridine orange or LysoTracker. Prolonged exposure of cells expressing ZnT-2 to zinc causes an accretion of intracellular vesicles. We suggest that ZnT-2 protects these cells from zinc toxicity by facilitating zinc transport into an endosomal/lysosomal compartment.
MeSH Terms
Amino Acid Sequence
Animals
Base Sequence
Biological Transport, Active
Carrier Proteins/genetics,metabolism
Cation Transport Proteins
Cell Line
Chromosome Mapping
Cricetinae
DNA, Complementary/genetics
Membrane Proteins/genetics,metabolism
Mice
Mice, Inbred C57BL
Microscopy, Fluorescence
Molecular Sequence Data
Muridae
Mutation
Rats
Sequence Homology, Amino Acid
Subcellular Fractions/metabolism
Transfection
Zinc/metabolism,toxicity
Chemicals
Carrier Proteins
Cation Transport Proteins
DNA, Complementary
Membrane Proteins
Slc30a1 protein, mouse
Slc30a2 protein, rat
Slc30a1 protein, rat
Zinc
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Palmiter R D
Howard Hughes Medical Institute, University of Washington, Seattle 98195-7370, USA.
Cole T B
Findley S D
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