We created a series of transgenic mice which either overexpress methionine sulfoxide reductase A in specific cellular locations or which lack the enzyme. We also established a mouse lacking methionine sulfoxide reductase B1 and successfully crossed it to create a double knockout mouse lacking both the A and B1 isozymes. The role of these reductases in resistance to oxidative stress during ischemic-reperfusion was studied with the Langendorff model. We expected that the transgenic mouse overexpressing methionine sulfoxide reductase A in mitochondria would be protected against injury while one or more of the knockout animals would experience more severe injury. This is not what we found. Rather, the knockout animals experienced the same degree of injury as the wild-type mice. Further, only the transgene expressing cytosolically targeted methionine sulfoxide reductase A was protected from ischemia-reperfusion induced cardiac injury. Importantly, the enzyme must be myristoylated to provide protection. A transgenic mouse overexpressing a non-myristolated methionine sulfoxide reductase A was not protected from injury. We are also completing an investigation of the hypothesis that methionine sulfoxide reductase is important in nutrition, functioning to salvage the metabolically """"""""expensive"""""""" amino acid methionine by reducing its sulfoxide. Weanling animals with different genotypes were raised on diets varying in methionine content, and their growth and biochemical parameters were being measured. This large body of data is now being analyzed. While methionine sulfoxide reductase A is well-known to reduce methionine sulfoxide to methionine, we have now demonstrated that it is capable of catalyzing the reverse reaction: methionine sulfoxide reductase A is also a methionine oxidase. The enzyme stereospecifically oxidizes free methionine, methionine in peptides, and methionine in proteins. Because the reaction is stereospecific, the enzyme can also fully reverse the reaction. This finding establishes a mechanistic basis for methionine sulfoxide reductase A to function in cellular regulation and signaling.

Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
National Heart, Lung, and Blood Institute
Zip Code
Williamson, Kim C; Levine, Rodney L; Miller, Louis H (2018) Even malaria parasites watch their host's diet. Nat Microbiol 3:130-131
Werner-Allen, Jonathan W; Monti, Sarah; DuMond, Jenna F et al. (2018) Isoindole Linkages Provide a Pathway for DOPAL-Mediated Cross-Linking of ?-Synuclein. Biochemistry 57:1462-1474
Lim, Jung Mi; Lim, Jung Chae; Kim, Geumsoo et al. (2018) Myristoylated methionine sulfoxide reductase A is a late endosomal protein. J Biol Chem 293:7355-7366
Lim, Jung Mi; Kim, Geumsoo; Levine, Rodney L (2018) Methionine in Proteins: It's Not Just for Protein Initiation Anymore. Neurochem Res :
Freeman, Lita A; Demosky Jr, Stephen J; Konaklieva, Monika et al. (2017) Lecithin:Cholesterol Acyltransferase Activation by Sulfhydryl-Reactive Small Molecules: Role of Cysteine-31. J Pharmacol Exp Ther 362:306-318
Werner-Allen, Jon W; Levine, Rodney L; Bax, Ad (2017) Superoxide is the critical driver of DOPAL autoxidation, lysyl adduct formation, and crosslinking of ?-synuclein. Biochem Biophys Res Commun 487:281-286
Uehara, Hiroshi; Luo, Shen; Aryal, Baikuntha et al. (2016) Distinct oxidative cleavage and modification of bovine [Cu- Zn]-SOD by an ascorbic acid/Cu(II) system: Identification of novel copper binding site on SOD molecule. Free Radic Biol Med 94:161-73
Werner-Allen, Jon W; DuMond, Jenna F; Levine, Rodney L et al. (2016) Toxic Dopamine Metabolite DOPAL Forms an Unexpected Dicatechol Pyrrole Adduct with Lysines of ?-Synuclein. Angew Chem Int Ed Engl 55:7374-8
Salmon, Adam B; Kim, Geumsoo; Liu, Chengyu et al. (2016) Effects of transgenic methionine sulfoxide reductase A (MsrA) expression on lifespan and age-dependent changes in metabolic function in mice. Redox Biol 10:251-256
Kim, Geumsoo; Levine, Rodney L (2016) A Methionine Residue Promotes Hyperoxidation of the Catalytic Cysteine of Mouse Methionine Sulfoxide Reductase A. Biochemistry 55:3586-93

Showing the most recent 10 out of 44 publications