title: The DinB superfamily includes novel mycothiol, bacillithiol, and glutathione S-transferases.
abstract: The superfamily of glutathione S-transferases has been the subject of extensive study; however, Actinobacteria produce mycothiol (MSH) in place of glutathione, and no mycothiol S-transferase (MST) has been identified. Using mycothiol and monochlorobimane as substrates, an MST activity was detected in extracts of Mycobacterium smegmatis and purified sufficiently to allow identification of MSMEG_0887, a member the DUF664 family of the DinB superfamily, as the MST. The identity of the M. smegmatis and homologous Mycobacterium tuberculosis (Rv0443) enzymes was confirmed by cloning, and the expressed proteins were found to be active with MSH but not bacillithiol (BSH) or glutathione (GSH). Bacillus subtilis YfiT is another member of the DinB superfamily, but this bacterium produces BSH. The YfiT protein was shown to have S-transferase activity with monochlorobimane when assayed with BSH but not with MSH or GSH. Enterococcus faecalis EF_3021 shares some homology with MSMEG_0887, but En. faecalis produces GSH but not MSH or BSH. Cloned and expressed EF_0321 was active with monochlorobimane and GSH but not with MSH or BSH. MDMPI_2 is another member of the DinB superfamily and has been previously shown to have mycothiol-dependent maleylpyruvate isomerase activity. Three of the eight families of the DinB superfamily include proteins shown to catalyze thiol-dependent metabolic or detoxification activities. Because more than two-thirds of the sequences assigned to the DinB superfamily are members of these families, it seems likely that such activity is dominant in the DinB superfamily.
authors: Department of Chemistry and Biochemistry, University of California at San Diego, La Jolla, California 92093, United States.
authors: Gerald L
authors: GL
authors: Newton
authors: Stephan S
authors: SS
authors: Leung
authors: Judy I
authors: JI
authors: Wakabayashi
authors: Mamta
authors: M
authors: Rawat
authors: Robert C
authors: RC
authors: Fahey
mesh_terms: D000581:Amidohydrolases / Q000737:chemistry / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D003001:Cloning, Molecular; D003545:Cysteine / Q000031:analogs & derivatives* / Q000378:metabolism; D013293:Enterococcus faecalis / Q000201:enzymology; D005944:Glucosamine / Q000031:analogs & derivatives* / Q000378:metabolism; D005982:Glutathione Transferase / Q000737:chemistry* / Q000378:metabolism*; D006020:Glycopeptides / Q000378:metabolism*; D007294:Inositol / Q000378:metabolism*; D005810:Multigene Family; D020102:Mycobacterium smegmatis / Q000378:metabolism; D009169:Mycobacterium tuberculosis / Q000201:enzymology; D010802:Phylogeny; D011720:Pyrazoles / Q000378:metabolism; D017386:Sequence Homology, Amino Acid
chemical_list: D001426:Bacterial Proteins; D006020:Glycopeptides; D011720:Pyrazoles; C543521:bacillithiol; C089265:mycothiol; D007294:Inositol; C059597:monochlorobimane; D005982:Glutathione Transferase; D000581:Amidohydrolases; C022427:polysaccharide deacetylase; D003545:Cysteine; D005944:Glucosamine