Methionine sulfoxide reductases (MsrA and MsrB) are repair enzymes that reduce methionine sulfoxide (MetO) residues in proteins to methionine (Met) in a stereospecific manner. These enzymes protect cells from oxidative stress and have been implicated in delaying the aging process and progression of neurodegenerative diseases. Mammals possess one MsrA and three MsrBs. The major mammalian MsrB, MsrB1, is a selenocysteine-containing protein, whose expression and activity can be regulated by dietary selenium. In this application, we propose to characterize regulation of protein function by stereospecific, site-specific, reversible Met oxidation;define the roles of Met, MetO and enzymes that act upon them in the regulation of lifespan;and establish a systems-level understanding of Met oxidation.
Specific Aim 1 is based on the finding that mouse Mical proteins specifically oxidize conserved Met residues in actin, which are then reduced back to Met by MsrB1. Further studies may add MetO to a list of posttranslational modifications, such as phosphorylation, acetylation, and methylation, that regulate protein function. We will characterize Mical-based oxidation of Met residues in actin and MsrB-based reduction of MetO residues, determine the redox state of Met in actin using unique reagents and examine physiological relevance of actin regulation through reversible Met oxidation.
Specific Aim 2 will address discrepancy in the regulation of lifespan by various Msrs. We will use transgenic fruit flies expressing various Msrs to test the hypothesis that the reduction of free MetO contributes to lifespan control by these enzymes. In addition, we will employ Met and MetO diets to test the hypothesis that the regulation of lifespan by Met depends on the status of other amino acids.
Specific Aim 3 will identify proteins with MetO by using advanced proteomic methods and characterize Met reactivity on the proteome scale. We will also employ stereospecific MetO sensors to characterize these species in cells and biological samples.

Public Health Relevance

Oxidative damage to proteins is considered to be one of the major factors that lead to or accompany aging. This damage results from oxidation of amino acids, among which methionine is notable because of its abundance and susceptibility to oxidation and repair. Methionine sulfoxide reductases are oxidoreductases that reduce the oxidized forms of methionine, methionine sulfoxides, in a stereospecific manner. We propose to address critical deficiencies in our understanding of control of protein function by reversible methionine oxidation, characterize regulation of lifespan by methionine sulfoxide reductases, methionine and methionine sulfoxide, and develop systems level understanding of methionine oxidation in cells and organisms.

Agency
National Institute of Health (NIH)
Type
Research Project (R01)
Project #
2R01AG021518-12
Application #
8631557
Study Section
Integrative Nutrition and Metabolic Processes Study Section (INMP)
Program Officer
Finkelstein, David B
Project Start
Project End
Budget Start
Budget End
Support Year
12
Fiscal Year
2014
Total Cost
Indirect Cost
Name
Brigham and Women's Hospital
Department
Type
DUNS #
City
Boston
State
MA
Country
United States
Zip Code
02115
Labunskyy, Vyacheslav M; Suzuki, Yo; Hanly, Timothy J et al. (2014) The insertion Green Monster (iGM) method for expression of multiple exogenous genes in yeast. G3 (Bethesda) 4:1183-91
Lee, Byung Cheon; Kaya, Alaattin; Ma, Siming et al. (2014) Methionine restriction extends lifespan of Drosophila melanogaster under conditions of low amino-acid status. Nat Commun 5:3592
Fang, Xiaodong; Seim, Inge; Huang, Zhiyong et al. (2014) Adaptations to a subterranean environment and longevity revealed by the analysis of mole rat genomes. Cell Rep 8:1354-64
Naranjo-Suarez, Salvador; Carlson, Bradley A; Tobe, Ryuta et al. (2013) Regulation of HIF-1? activity by overexpression of thioredoxin is independent of thioredoxin reductase status. Mol Cells 36:151-7
Le, Dung Tien; Tarrago, Lionel; Watanabe, Yasuko et al. (2013) Diversity of plant methionine sulfoxide reductases B and evolution of a form specific for free methionine sulfoxide. PLoS One 8:e65637
Seim, Inge; Fang, Xiaodong; Xiong, Zhiqiang et al. (2013) Genome analysis reveals insights into physiology and longevity of the Brandt's bat Myotis brandtii. Nat Commun 4:2212
Lee, Byung Cheon; Peterfi, Zalan; Hoffmann, Fukun W et al. (2013) MsrB1 and MICALs regulate actin assembly and macrophage function via reversible stereoselective methionine oxidation. Mol Cell 51:397-404
Gladyshev, Vadim N (2013) The origin of aging: imperfectness-driven non-random damage defines the aging process and control of lifespan. Trends Genet 29:506-12
Kasaikina, Marina V; Hatfield, Dolph L; Gladyshev, Vadim N (2012) Understanding selenoprotein function and regulation through the use of rodent models. Biochim Biophys Acta 1823:1633-42
Labunskyy, Vyacheslav M; Lee, Byung Cheon; Handy, Diane E et al. (2011) Both maximal expression of selenoproteins and selenoprotein deficiency can promote development of type 2 diabetes-like phenotype in mice. Antioxid Redox Signal 14:2327-36

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