Presence of a metallothionein-like protein in the bovine pineal gland.
The high concentration of zinc in the bovine pineal gland prompted us to investigate the existence of a zinc-binding protein in this organ. In this study, we report that the subcellular distribution of zinc in the bovine pineal gland is nonuniform, with the crude nuclear, mitochondrial, microsomal, and supernatant fractions having 0.264 +/- 0.038, 0.160 +/- 0.019, 0.130 +/- 0.016, and 0.287 +/- 0.010 micrograms zinc/mg protein, respectively. Furthermore, gel filtration studies using Sephadex G-75 and a 105,000 g supernatant fraction revealed two zinc binding protein peaks that bind 1.7 and 3.7 micrograms Zn++/mg protein, respectively. Furthermore, purification of the protein peak with an elution volume (ve/vo) of 2.06 on anion exchange chromatography (DEAE-A25) yielded a single protein peak which binds 10 micrograms zinc/mg protein. The comparative high performance liquid chromatographic (HPLC) profiles of the zinc-induced hepatic metallothionein isoform I (retention time = 17.39 min) and of the bovine pineal metallothionein-like protein isoform I (retention time = 17.49 min) are similar. Since zinc is a potent inhibitor of sulfhydryl-containing enzymes and receptor sites, we investigated the effects of zinc and found that it inhibited the binding of [3H]glutamate (IC 50 = 80 microM) and of [3H]spiroperidol (IC 50 = 0.6 mM) to the pineal membranes. The results of these studies are interpreted to indicate that the bovine pineal gland possesses an active and dynamic zinc homeostatic mechanism, whose precise function(s) remain(s) to be delineated.
Awad
A
A
Department of Pharmacology, University of Nebraska College of Medicine, Omaha.
Govitrapong
P
P
Hama
Y
Y
Hegazy
M
M
Ebadi
M
M
D000818:Animals; D002352:Carrier Proteins / Q000302:isolation & purification*; D002417:Cattle; D066298:In Vitro Techniques; D008297:Male; D008668:Metallothionein / Q000302:isolation & purification*; D010870:Pineal Gland / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains; D011954:Receptors, Dopamine / Q000187:drug effects / Q000378:metabolism; D017470:Receptors, Glutamate; D017981:Receptors, Neurotransmitter / Q000187:drug effects / Q000378:metabolism; D013134:Spiperone / Q000378:metabolism; D015032:Zinc / Q000378:metabolism*
D002352:Carrier Proteins; D011954:Receptors, Dopamine; D017470:Receptors, Glutamate; D017981:Receptors, Neurotransmitter; D013134:Spiperone; D008668:Metallothionein; D015032:Zinc