Expression, subcellular localization, and antioxidant role of mammalian methionine sulfoxide reductases in Saccharomyces cerevisiae

Abstract

Despite the growing body of evidence suggesting a role for MsrA in antioxidant defense, little is currently known regarding the function of MsrB in cellular protection against oxidative stress. In this study, we overexpressed the mammalian MsrB and MsrA genes in Saccharomyces cerevisiae and assessed their subcellular localization and antioxidant functions. We found that the mitochondrial MsrB3 protein (MsrB3B)

Methionine Sulfoxide Reductase A, B1 and B2 Are Likely to Be Involved in the Protection against Oxidative Stress in the Inner Ear

“…Next, the tissue sections were blocked with 5% normal goat serum in PBS for 1 h at room temperature, followed by incubation with each primary antibody overnight at 4 ° C [antibody dilutions: MsrA, 1: 100; MsrB1, 1: 100; MsrB2, 1: 50; myosin VIIA (MYO7A) 1: 500; neurofilament-200 (NF200), 1: 500]. Each primary antibody of the Msr family was validated in previous studies [Kim and Gladyshev, 2004b;Kwak et al, 2009]. The tissue sections were incubated with secondary antibody, either a goat anti-rabbit IgG antibody conjugated with Alexa 555 fluorescent or a goat anti-mouse IgG antibody conjugated with Alexa 488 fluorescent (Molecular Probes/Invitrogen, Life Technology, Carlsbad, Calif., USA), at 1: 1,000 dilution for 1 h at room temperature and then stained with 4 ′ -6-diamidino-2-phenylindole (Roche Diagnostics, Indianapolis, Ind., USA) for 5 min at room temperature and visualized using a confocal laser scanning microscope (LSM700; Zeiss, Oberkochen, Germany).…”

Dimethyl sulfoxide elevates hydrogen peroxide-mediated cell death in Saccharomyces cerevisiae by inhibiting the antioxidant function of methionine sulfoxide reductase A

“…Deletion or knockdown of MsrA and MsrB genes in organisms ranging from bacteria to mammals results in increased sensitivity to oxidative stress (17,(19)(20)(21)(22)(23). Conversely, overexpression of these genes protects against oxidative stress-induced cell death (24)(25)(26)(27). In addition, fRMsr also has an antioxidant function in yeast (15).…”

Genes and Hearing Loss: Relationship to Oxidative Stress and Free Radical Formation

“…MSRB3 encodes a protein with methionine sulfoxide reductase activity (Weissbach et al 2002 ) and is localized to sensory and supporting cells in the organ of Corti (Ahmed et al 2011 ;Kwon et al 2014 ). While studies in cultured cells are consistent with an infl uence on ER and oxidative stress resistance (Kwak et al 2009(Kwak et al , 2012, the lack of an overt accumulation of aberrantly oxidized proteins in the cochlea of mice carrying a targeted deletion of MSRB3 suggests that a more subtle effect on redox status may underlie the stereocilia and hearing defects in these mutants (Kwon et al 2014 ).…”