title: Adaptation of microsomal glutathione transferase 1 in PC12 cells with modified PMCA isoforms composition.
abstract: Microsomal glutathione transferase 1 (MGST1) is an integral homo-trimeric membrane protein with transferase and peroxidase activities. With glutathione as a co-substrate, it scavenges toxic compounds and may exert anti-apoptotic effect. We examined the effect of suppression of plasma membrane Ca(2+)-ATPase isoforms--PMCA2 or PMCA3 on MGST1 in PC12 cells. GSH level was significantly higher in PMCA2-reduced line, but similar GSSG/GSH ratios in all cell lines suggested an efficient protection or absence of oxidative stress. The ATP concentration decreased in both modified lines, although in PMCA2-suppressed cells the decrease was higher. Total GSTs activity in postmitochondrial fraction increased by 30% in the cells with reduced PMCA3. After treatment with MGST1 activator N-ethylmaleimide (NEM), the activity increased in both transfected lines by 30-40%. Real-time PCR also showed a higher mRNA expression of MGST1 in these lines. Staining with antibody recognizing all cytosolic and membrane-bound GSTs revealed the difference in oligomeric forms of GSTs, and specific anti-MGST1 antibody showed the presence of MGST1 hexamers in the transfected cells. Formation of similar hexamers was detected in the control line after treatment with peroxynitrite. Modification of MGST1 under reduced PMCAs amount may represent an adaptive mechanism that offers protection against the cytotoxicity mediated by increased Ca2+.
authors.affiliation: Department of Molecular Neurochemistry, Medical University of Lodz, 6/8 Mazowiecka Street, 92-215, Lodz, Poland.
authors.forename: Tomasz
authors.initials: T
authors.lastname: Boczek
authors.forename: Anna
authors.initials: A
authors.lastname: Kozaczuk
authors.forename: Joanna
authors.initials: J
authors.lastname: Taha
authors.forename: Bozena
authors.initials: B
authors.lastname: Ferenc
authors.forename: Ludmila
authors.initials: L
authors.lastname: Zylinska
mesh_terms: D000222:Adaptation, Physiological / Q000502:physiology; D000818:Animals; D004789:Enzyme Activation; D005982:Glutathione Transferase / Q000378:metabolism*; D008861:Microsomes / Q000201:enzymology*; D016716:PC12 Cells; D053499:Plasma Membrane Calcium-Transporting ATPases / Q000378:metabolism*; D051381:Rats
chemical_list: C122107:microsomal glutathione S-transferase-I; D005982:Glutathione Transferase; D053499:Plasma Membrane Calcium-Transporting ATPases