Identification of metallothionein receptors in human astrocytes.
Metallothionein (MT) isoforms are low molecular weight (6000-7000 Da) zinc binding proteins containing 60-68 amino acid residues, 25-30% cysteine, no aromatic amino acids, and binding between 5-7 g zinc/mol of protein. Since the synthesis of MT is induced by endotoxin, cytokines, and glucocorticoids, MT is now considered to be an acute phase protein protecting against oxygen radicals and oxidative damages caused by inflammation, tissue injury, and stress to the central nervous system. By postulating that a specific mechanism must exist to foster the induction of MTs I and II by numerous and diversified factors, we searched for and identified for the first time, MT receptors on U373MG cell membrane preparations, by using fluoresceinated MT I isoform probe; and by employing cysteine, glutathione, and four MT isoforms to determine high affinity and specific binding. MT receptors revealed a Kd value of 0.84 nM and a Bmax of 99.82 fmol/mg protein. Moreover, MT receptors were found in greater density on the surface of aggregated astrocytes. We postulate that conditions or agents generating reactive oxygen species may influence the expression of MT receptors.
El Refaey
H
H
Department of Pharmacology and Pathology, University of Nebraska College of Medicine, Omaha 68198-6260, USA.
Ebadi
M
M
Kuszynski
C A
CA
Sweeney
J
J
Hamada
F M
FM
Hamed
A
A
D000595:Amino Acid Sequence; D000704:Analysis of Variance; D001253:Astrocytes / Q000378:metabolism*; D002478:Cells, Cultured; D003545:Cysteine / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D016650:Fluorescein-5-isothiocyanate / Q000032:analysis; D005456:Fluorescent Dyes; D005978:Glutathione / Q000494:pharmacology; D006801:Humans; D008668:Metallothionein / Q000031:analogs & derivatives / Q000378:metabolism*; D008969:Molecular Sequence Data; D011956:Receptors, Cell Surface / Q000378:metabolism*; D013997:Time Factors; D015032:Zinc / Q000032:analysis
D005456:Fluorescent Dyes; D011956:Receptors, Cell Surface; C109611:metallothionein receptor; D008668:Metallothionein; D005978:Glutathione; D016650:Fluorescein-5-isothiocyanate; D015032:Zinc; D003545:Cysteine