title: Efflux permease CgAcr3-1 of Corynebacterium glutamicum is an arsenite-specific antiporter.
abstract: Resistance to arsenite (As(III)) by cells is generally accomplished by arsenite efflux permeases from Acr3 or ArsB unrelated families. We analyzed the function of three Acr3 proteins from Corynebacterium glutamicum, CgAcr3-1, CgAcr3-2, and CgAcr3-3. CgAcr3-1 conferred the highest level of As(III) resistance and accumulation in vivo. CgAcr3-1 was also the most active when everted membranes vesicles from Escherichia coli or C. glutamicum mutants were assayed for efflux with different energy sources. As(III) and antimonite (Sb(III)) resistance and accumulation studies using E. coli or C. glutamicum arsenite permease mutants clearly show that CgAcr3-1 is specific for As(III). In everted membrane vesicles expressing CgAcr3-1, dissipation of either the membrane potential or the pH gradient of the proton motive force did not prevent As(III) uptake, whereas dissipation of both components eliminated uptake. Further, a mutagenesis study of CgAcr3-1 suggested that a conserved cysteine and glutamate are involved in active transport. Therefore, we propose that CgAcr3-1 is an antiporter that catalyzes arsenite-proton exchange with residues Cys129 and Glu305 involved in efflux.
authors.affiliation: Departamento de Biología Molecular, Área de Microbiología, Facultad de Biología-Ambientales, Universidad de León, 24071 León, Spain.
authors.forename: Almudena F
authors.initials: AF
authors.lastname: Villadangos
authors.affiliation: Department of Biochemistry and Molecular Biology, Wayne State University School of Medicine, Detroit, Michigan 48201.
authors.forename: Hsueh-Liang
authors.initials: HL
authors.lastname: Fu
authors.affiliation: Departamento de Biología Molecular, Área de Microbiología, Facultad de Biología-Ambientales, Universidad de León, 24071 León, Spain.
authors.forename: Jose A
authors.initials: JA
authors.lastname: Gil
authors.affiliation: Department of Structural Biology, Vlaams Instituut voor Biotechnologie; Structural Biology Brussels, Vrije Universiteit Brussels, 1050 Brussels, Belgium; Brussels Center for Redox Biology, 1050 Brussels, Belgium.
authors.forename: Joris
authors.initials: J
authors.lastname: Messens
authors.affiliation: Department of Cellular Biology and Pharmacology, Herbert Wertheim College of Medicine, Florida International University, Miami, Florida 33099. Electronic address: brosen@fiu.edu.
authors.forename: Barry P
authors.initials: BP
authors.lastname: Rosen
authors.affiliation: Departamento de Biología Molecular, Área de Microbiología, Facultad de Biología-Ambientales, Universidad de León, 24071 León, Spain. Electronic address: luis.mateos@unileon.es.
authors.forename: Luis M
authors.initials: LM
authors.lastname: Mateos
mesh_terms: D000595:Amino Acid Sequence; D017920:Antiporters / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D018053:Arsenites / Q000378:metabolism* / Q000633:toxicity; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D055162:Biocatalysis; D001692:Biological Transport; D002462:Cell Membrane / Q000378:metabolism; D048230:Corynebacterium glutamicum / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D057927:Hydrophobic and Hydrophilic Interactions; D008564:Membrane Potentials; D026901:Membrane Transport Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D017434:Protein Structure, Tertiary; D011522:Protons; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity
chemical_list: D017920:Antiporters; D018053:Arsenites; D001426:Bacterial Proteins; D026901:Membrane Transport Proteins; D011522:Protons; C015001:arsenite
references.citation: Biochim Biophys Acta. 2011 Jul;1808(7):1855-9
references.pmid: 21447319
references.citation: Biometals. 2009 Feb;22(1):117-30
references.pmid: 19130261
references.citation: Proc Natl Acad Sci U S A. 1999 Apr 27;96(9):5001-6
references.pmid: 10220408
references.citation: Appl Microbiol Biotechnol. 1996 Dec;46(5-6):554-8
references.pmid: 9008889
references.citation: J Biol Chem. 2006 Apr 14;281(15):9925-34
references.pmid: 16467301
references.citation: FEBS J. 2007 Feb;274(3):612-29
references.pmid: 17288550
references.citation: J Biol Chem. 1990 Jan 5;265(1):190-4
references.pmid: 1688427
references.citation: J Biol Chem. 2001 Jul 13;276(28):26301-7
references.pmid: 11306588
references.citation: Res Microbiol. 2007 Mar;158(2):128-37
references.pmid: 17258434
references.citation: FEBS Lett. 2010 Jul 16;584(14):3089-94
references.pmid: 20553716
references.citation: Adv Exp Med Biol. 2010;679:47-55
references.pmid: 20666223
references.citation: Int Microbiol. 2006 Sep;9(3):207-15
references.pmid: 17061211
references.citation: Biochim Biophys Acta. 2010 Nov;1798(11):2170-5
references.pmid: 20655873
references.citation: Nature. 1970 Aug 15;227(5259):680-5
references.pmid: 5432063
references.citation: J Biol Chem. 2004 Apr 30;279(18):18334-41
references.pmid: 14970228
references.citation: J Bacteriol. 2006 Jan;188(2):409-23
references.pmid: 16385030
references.citation: Trends Biochem Sci. 1993 Jan;18(1):13-20
references.pmid: 8438231
references.citation: Plant Cell. 2010 Jun;22(6):2045-57
references.pmid: 20530755
references.citation: J Bacteriol. 1996 Oct;178(19):5768-75
references.pmid: 8824624
references.citation: Biochimie. 2009 Oct;91(10):1229-37
references.pmid: 19567262
references.citation: Mol Microbiol. 2011 Feb;79(4):872-81
references.pmid: 21299644
references.citation: Physiol Rev. 2006 Jul;86(3):849-99
references.pmid: 16816140
references.citation: J Biol Chem. 2009 Jul 24;284(30):19887-95
references.pmid: 19494117
references.citation: Mol Membr Biol. 2008 Dec;25(8):691-705
references.pmid: 19039703
references.citation: FEMS Yeast Res. 2004 Sep;4(8):821-32
references.pmid: 15450189
references.citation: Appl Environ Microbiol. 2005 Oct;71(10):6206-15
references.pmid: 16204540
references.citation: J Biol Chem. 1992 Jun 25;267(18):12570-6
references.pmid: 1535622
references.citation: J Biol Chem. 1986 Jan 15;261(2):678-83
references.pmid: 3001076
references.citation: J Biol Chem. 2008 Sep 12;283(37):25706-25714
references.pmid: 18591244
references.citation: J Bacteriol. 1998 Apr;180(7):1655-61
references.pmid: 9537360
references.citation: J Bacteriol. 2003 Sep;185(18):5363-71
references.pmid: 12949088
references.citation: J Biol Chem. 1994 Oct 14;269(41):25442-6
references.pmid: 7929243
references.citation: Proc Natl Acad Sci U S A. 1997 Sep 16;94(19):10128-31
references.pmid: 9294174
references.citation: J Biol Chem. 1986 Nov 15;261(32):15030-8
references.pmid: 3021763
references.citation: J Biol Chem. 1997 Jan 3;272(1):326-31
references.pmid: 8995265