Abstract
DNA-binding proteins from nutrient-starved cells (DPS) protect cells from oxidative stress by removing H2O2 and iron. A new class of DPS-like proteins has recently been identified, with DPS-like protein from Sulfolobus solfataricus (SsDPS) being the best characterized to date. SsDPS protects cells from oxidative stress and is upregulated in response to H2O2 but also in response to iron depletion. The ferroxidase active site of SsDPS is structurally similar to the active sites of manganese catalase and rat liver arginase. The present work shows that the ferroxidase center in SsDPS binds two Mn2+ ions with K D = (1/K 1 K 2)1/2 = 48(3) μM. The binding constant of the second Mn2+ is significantly higher than that of the first, inducing dinuclear Mn(II) cluster formation for all but the lowest concentrations of added Mn2+. In competition experiments, equimolar amounts of Fe2+ were unable to displace the bound manganese. EPR spectroscopy of the Mn2 2+ cluster showed signals comparable to those of other characterized dimanganese clusters. The exchange coupling for the cluster was determined, J = −1.4(3) cm−1 (H = −2JS 1 S 2), and is within the range expected for a μ1,1-carboxylato bridge between the manganese ions. Manganese-bound SsDPS showed catalase activity at a rate 10–100 times slower than for manganese catalases. EPR spectra of SsDPS after addition of H2O2 showed the appearance of an intermediate in the reaction with H2O2.
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Abbreviations
- DPS:
-
DNA-binding protein from nutrient-starved cells
- EPR:
-
Electron paramagnetic resonance
- MnCat:
-
Manganese catalase
- ROS:
-
Reactive oxygen species
- SsDPS:
-
DNA-binding protein from nutrient-starved cells like protein from Sulfolobus solfataricus
References
Fang FC (2004) Nat Rev Microbiol 2:820–832
Halsey TA, Vazquez-Torres A, Gravdahl DJ, Fang FC, Libby SJ (2004) Infect Immun 72:1155–1158
Li X, Pal U, Ramamoorthi N, Liu X, Desrosiers DC, Eggers CH, Anderson JF, Radolf JD, Fikrig E (2007) Mol Microbiol 63:694–710
Grant RA, Filman DJ, Finkel SE, Kolter R, Hogle JM (1998) Nat Struct Biol 5:294–303
Smith JL (2004) Crit Rev Microbiol 30:173–185
Zhao G, Ceci P, Ilari A, Giangiacomo L, Laue TM, Chiancone E, Chasteen ND (2002) J Biol Chem 277:27689–27696
Lawson TL, Crow A, Lewin A, Yasmin S, Moore GR, Brun NEL (2009) Biochemistry 48:9031–9039
Crow A, Lawson TL, Lewin A, Moore GR, Brun NEL (2009) J Am Chem Soc 131:6808–6813
Wiedenheft B, Mosolf J, Willits D, Yeager M, Dryden KA, Young M, Douglas T (2005) Proc Natl Acad Sci USA 102:10551–10556
Gauss GH, Benas P, Wiedenheft B, Young M, Douglas T, Lawrence CM (2006) Biochemistry 45:10815–10827
Barynin VV, Whittaker MM, Antonyuk SV, Lamzin VS, Harrison PM, Artymiuk PJ, Whittaker JW (2001) Structure (Camb) 9:725–738
Cama E, Emig FA, Ash DE, Christianson DW (2003) Biochemistry 42:7748–7758
Golynskiy MV, Gunderson WA, Hendrich MP, Cohen SM (2006) Biochemistry 45:15359–15372
Pierce BS, Elgren TE, Hendrich MP (2003) J Am Chem Soc 125:8748–8759
Golombek AP, Hendrich MP (2003) J Magn Reson 165:33–48
Gunderson WA, Zatsman AI, Emerson JP, Farquhar ER, Que L Jr, Lipscomb JD, Hendrich MP (2008) J Am Chem Soc 130:14465–14467
Martell AE, Smith RM (eds) (1974) Critical stability constants, vol 1: amino acids. Plenum Press, New York, p 469
Khangulov SV, Barynin VV, Antonyuk-Barynina SV (1990) Biochim Biophys Acta 1020:25–33
Allgood GS, Perry JJ (1986) J Bacteriol 168:563–568
Shank M, Barynin V, Dismukes GC (1994) Biochemistry 33:15433–15436
Meier AE, Whittaker MM, Whittaker JW (1996) Biochemistry 35:348–360
Khangulov SV, Pessiki PJ, Barynin VV, Ash DE, Dismukes GC (1995) Biochemistry 34:2015–2025
Smoukov SK, Telser J, Bernat BA, Rife CL, Armstrong RN, Hoffman BM (2002) J Am Chem Soc 124:2318–2326
Boelrijk AE, Dismukes GC (2000) Inorg Chem 39:3020–3028
Wu WAJ, Penner-Hahn JE, Pecoraro VL (2004) Chem Rev 104:903–938
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Hayden, J.A., Hendrich, M.P. EPR spectroscopy and catalase activity of manganese-bound DNA-binding protein from nutrient starved cells. J Biol Inorg Chem 15, 729–736 (2010). https://doi.org/10.1007/s00775-010-0640-3
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DOI: https://doi.org/10.1007/s00775-010-0640-3