JBIC Journal of Biological Inorganic Chemistry

, Volume 19, Issue 6, pp 1009–1024 | Cite as

Electrostatic effects control the stability and iron release kinetics of ovotransferrin

  • Sandeep Kumar
  • Deepak Sharma
  • Rajesh Kumar
  • Rajesh Kumar
Original Paper

Abstract

The contribution of electrostatic interactions to the stability of ovotransferrin-Fe3+ (oTf-Fe3+) complex has been assessed by equilibrium experiments that measure iron retention level of diferric-ovotransferrin (Fe2oTf) as a function of pH and urea in the presence of salts (NaCl, Na2SO4, NaBr, NaNO3) and sucrose at 25 °C. As [salt] is increased, the pH-midpoint for iron release increases monoexponentially and plateau at ~0.4(±0.05) M NaCl/NaBr/NaNO3 or ~0.15(±0.03) M Na2SO4. However, at pH 7.4, the urea-midpoints for iron release (based on fluorescence emission at 340 nm) and for unfolding of Fe2oTf and apo-ovotransferrin (based on ellipticity values at 222 and 282 nm) decrease at low salt concentrations [≤0.1(±0.02) M Na2SO4 or ≤0.35(±0.15) M NaCl], but increase at higher salt concentrations. Furthermore, Na2SO4 has a greater effect than NaCl in increasing the urea-midpoints for iron release and unfolding. These results indicate that at low salt concentrations, the electrostatic effects destabilize the oTf-Fe3+ complex and also decrease the structural stability of the proteins. In contrast, at higher concentrations, salt ions behave according to Hofmeister series. At pH 5.6, as [salt] is increased, the rate constants for reductive iron release (Fe2+ release) and urea denaturation-induced iron release (Fe3+ release) from the N-lobe of oTf (FeNoTf) increase monoexponentially and plateau at ~0.4(±0.1) M NaNO3/NaCl or ~0.2(±0.05) M Na2SO4. These results suggest that the anion-binding-induced conformational change as well as the electrostatic screening of surface Coulombic interactions plays important role in accelerating the iron release from FeNoTf under endosomal pH conditions.

Keywords

Iron release Electrostatic interactions Anion-binding Conformational change Structural stability 

Notes

Acknowledgments

This work was supported by DST-SERC Fast Track Research Grant (to R.K., project No. SR/FT/CS-070/2009), a Ramalingaswami Re-entry Fellowship by DBT, India (to D.S., BT/RLF/RE-ENTRY-33-2010), and UGC major grant (to R.K., F. No. 41-258/2012 (SR), Government of India.

Supplementary material

775_2014_1145_MOESM1_ESM.pdf (57 kb)
Supplementary material 1 (PDF 57 kb)

References

  1. 1.
    Jaenicke R, Bohm G (1998) Curr Opin Struct Biol 8:738–748PubMedGoogle Scholar
  2. 2.
    Pace CN (1995) Methods Enzymol 259:538–554PubMedGoogle Scholar
  3. 3.
    Karshikoff A, Ladenstein R (2007) In: Uversky VN, Permyakov E (eds) Nova Biomedical Books, New YorkGoogle Scholar
  4. 4.
    Perl D, Holtermann G, Schmid FX (2001) Biochemistry 40:15501–15511PubMedGoogle Scholar
  5. 5.
    Jayaraman S, Gantz DL, Gursky O (2006) Biochemistry 45:4620–4628PubMedGoogle Scholar
  6. 6.
    Benjwal S, Jayaraman S, Gursky O (2005) Biochemistry 44:10218–10226PubMedGoogle Scholar
  7. 7.
    Dominy BN, Perl D, Schmid FX, Brooks CL 3rd (2002) J Mol Biol 319:541–554PubMedGoogle Scholar
  8. 8.
    Vogt G, Woell S, Argos P (1997) J Mol Biol 269:631–643PubMedGoogle Scholar
  9. 9.
    Elcock AH, McCammon JA (1998) J Mol Biol 280:731–748PubMedGoogle Scholar
  10. 10.
    Kohn WD, Kay CM, Hodges RS (1997) J Mol Biol 267:1039–1052PubMedGoogle Scholar
  11. 11.
    Lee DA, Goodfellow JM (1998) Biophys J 85:2747–2759Google Scholar
  12. 12.
    Rinaldo D, Field MJ (2003) Biophys J 85:3485–3501PubMedCentralPubMedGoogle Scholar
  13. 13.
    Bobst CE, Zhang M, Kaltashov IA (2009) J Mol Biol 388:954–967PubMedCentralPubMedGoogle Scholar
  14. 14.
    Aisen P, Listowsky I (1980) Annu Rev Biochem 49:357–393PubMedGoogle Scholar
  15. 15.
    Dewan JC, Mikami B, Hirose M, Sacchettini JC (1993) Biochemistry 32:11963–11968PubMedGoogle Scholar
  16. 16.
    MacGillivray RT, Moore SA, Chen J, Anderson BF, Baker H, Luo Y, Bewley M, Smith CA, Murphy ME, Wang Y, Mason AB, Woodworth RC, Brayer GD, Baker EN (1998) Biochemistry 37:7919–7928PubMedGoogle Scholar
  17. 17.
    He QY, Mason AB, Tam BM, MacGillivray RT, Woodworth RC (1999) Biochemistry 38:9704–9711PubMedGoogle Scholar
  18. 18.
    Byrne SL, Chasteen ND, Steere AN, Mason AB (2010) J Mol Biol 396:130–140PubMedCentralPubMedGoogle Scholar
  19. 19.
    Steere AN, Byrne SL, Chasteen ND, Smith VC, MacGillivray RT, Mason AB (2010) J Biol Inorg Chem 15:1341–1352PubMedGoogle Scholar
  20. 20.
    James NG, Byrne SL, Steere AN, Smith VC, MacGillivray RT, Mason AB (2009) Biochemistry 48:2858–2867PubMedCentralPubMedGoogle Scholar
  21. 21.
    James NG, Berger CL, Byrne SL, Smith VC, MacGillivray RT, Mason AB (2007) Biochemistry 46:10603–10611PubMedGoogle Scholar
  22. 22.
    Halbrooks PJ, Giannetti AM, Klein JS, Björkman PJ, Larouche JR, Smith VC, MacGillivray RT, Everse SJ, Mason AB (2005) Biochemistry 44:15451–15460PubMedGoogle Scholar
  23. 23.
    Nurizzo D, Baker HM, He QY, MacGillivray RT, Mason AB, Woodworth RC, Baker EN (2001) Biochemistry 40:1616–1623PubMedGoogle Scholar
  24. 24.
    He QY, Mason AB, Tam BM, MacGillivray RT, Woodworth RC (1999) Biochem J 344:881–887PubMedCentralPubMedGoogle Scholar
  25. 25.
    Byrne SL, Mason AB (2009) J Biol Inorg Chem 14:771–781PubMedCentralPubMedGoogle Scholar
  26. 26.
    Spencer DS, Xu K, Logan TM, Zhou HX (2005) J Mol Biol 351:219–232PubMedGoogle Scholar
  27. 27.
    Hofmeister F (1888) Arch Exp Pathol Pharmakol 24:247–260Google Scholar
  28. 28.
    Baldwin RL (1996) Biophys J 71:2056–2063PubMedCentralPubMedGoogle Scholar
  29. 29.
    Perez-Jimenez R, Godoy-Ruiz R, Ibarra-Molero B, Sanchez-Ruiz JM (2004) Biophys J 86:2414–2429PubMedCentralPubMedGoogle Scholar
  30. 30.
    Kumar R, Mauk AG (2009) J Phys Chem B 113:12400–12409PubMedGoogle Scholar
  31. 31.
    Cacace MG, Landau EM, Ramsden JJ (1997) Q Rev Biophys 30:241–277PubMedGoogle Scholar
  32. 32.
    Von Hippel PH, Wong KY (1964) Science 145:577–580Google Scholar
  33. 33.
    Pegram LM, Record MT Jr (2008) J Phys Chem B 112:9428–9436PubMedCentralPubMedGoogle Scholar
  34. 34.
    Record MT Jr, Anderson CF, Lohman TM (1978) Q Rev Biophys 11:103–178PubMedGoogle Scholar
  35. 35.
    Villa A, Zecca L, Fusi P, Colombo S, Tedeschi G, Tortora P (1993) Biochem J 295:827–831PubMedCentralPubMedGoogle Scholar
  36. 36.
    Apetri AC, Surewicz WK (2003) J Biol Chem 278:22187–22192PubMedGoogle Scholar
  37. 37.
    Aasa R, Malmstrom BG, Saltman P, Vanngård T (1963) Biochim Biophys Acta 75:203–222PubMedGoogle Scholar
  38. 38.
    Sun H, Li H, Sadler PJ (1999) Chem Rev 99:2817–2842PubMedGoogle Scholar
  39. 39.
    MacGillivray RTA, Mason AB (2002) In: Transferrin Templeton DM (ed). Marcel Dekker, Inc., New YorkGoogle Scholar
  40. 40.
    He QY, Mason AB (2002) In: Templeton DM (ed). Marcel Dekker, Inc., New YorkGoogle Scholar
  41. 41.
    Guo M, Sun H, McArdle HJ, Gambling L, Sadler PJ (2000) Biochemistry 39:10023–10033PubMedGoogle Scholar
  42. 42.
    Tinoco AD, Valentine AM (2005) J Am Chem Soc 127:11218–11219PubMedGoogle Scholar
  43. 43.
    Tinoco AD, Incarvito CD, Valentine AM (2007) J Am Chem Soc 129:3444–3454PubMedGoogle Scholar
  44. 44.
    Hémadi M, Ha-Duong NT, Plantevin S, Vidaud C, El Hage Chahine JM (2010) J Biol Inorg Chem 15:497–504PubMedGoogle Scholar
  45. 45.
    Chikh Z, Ha-Duong NT, Miquel G, El Hage Chahine JM (2007) J Biol Inorg Chem 12:90–100PubMedGoogle Scholar
  46. 46.
    Zhong W, Parkinson JA, Guo M, Sadler PJ (2002) J Biol Inorg Chem 7:589–599PubMedGoogle Scholar
  47. 47.
    Dautry-Varsat A, Ciechanover A, Lodish HF (1983) Proc Natl Acad Sci USA 80:2258–2262PubMedCentralPubMedGoogle Scholar
  48. 48.
    Baker EN (1994) Adv Inorg Chem 41:389–463Google Scholar
  49. 49.
    Mason AB, Woodworth RC (1984) J Biol Chem 259:1866–1873Google Scholar
  50. 50.
    Mason AB, Brown SA, Church WR (1987) J Biol Chem 262:9011–9015PubMedGoogle Scholar
  51. 51.
    Mason AB, Woodworth RC, Oliver RW, Green BN, Lin LN, Brandts JF, Savage KJ, Tam BM, MacGillivray RT (1996) Biochem J 319:361–368PubMedCentralPubMedGoogle Scholar
  52. 52.
    Thibodeau SN, Lee DC, Palmiter RD (1978) J Biol Chem 253:3771–3774PubMedGoogle Scholar
  53. 53.
    Anderson BF, Baker HM, Norris GE, Rice DW, Baker EN (1989) J Mol Biol 209:711–734PubMedGoogle Scholar
  54. 54.
    Baker EN, Lindley PF (1992) J Inorg Biochem 47:147–160PubMedGoogle Scholar
  55. 55.
    Haridas M, Anderson BF, Baker EN (1995) Acta Crystallogr D Biol Crystallogr 51:629–646PubMedGoogle Scholar
  56. 56.
    Anderson BF, Baker HM, Norris GE, Rumball SV, Baker EN (1990) Nature 344:784–787PubMedGoogle Scholar
  57. 57.
    Kurokawa H, Mikami B, Hirose M (1995) J Mol Biol 254:196–207PubMedGoogle Scholar
  58. 58.
    Kurokawa H, Dewan JC, Mikami B, Sacchettini J, Hirose M (1999) J Biol Chem 274:28445–28452PubMedGoogle Scholar
  59. 59.
    Jeffrey PD, Bewley MC, MacGillivray RT, Mason AB, Woodworth RC, Baker EN (1998) Biochemistry 37:13978–13996PubMedGoogle Scholar
  60. 60.
    Sharma AK, Rajashankar KR, Yadav MP, Singh TP (1999) Acta Crystallogr Sect D55:1152–1157Google Scholar
  61. 61.
    Thorstensen K, Romslo I (1990) Biochem J 271:1–9PubMedCentralPubMedGoogle Scholar
  62. 62.
    Hemadi M, Ha-Duong NT, El HageChahine JM (2006) J Mol Biol 358:1125–1136PubMedGoogle Scholar
  63. 63.
    Schlabach MR, Bates GW (1975) J Biol Chem 25:2182–2188Google Scholar
  64. 64.
    Campbell RF, Chasteen ND (1977) J Biol Chem 252:5996–6001PubMedGoogle Scholar
  65. 65.
    Zweier JL, Wooten JB, Cohen JS (1981) Biochemistry 20:3505–3510PubMedGoogle Scholar
  66. 66.
    Aisen P, Leibma A, Pinkowitz RA, Pollack S (1973) Biochemistry 12:3679–3684PubMedGoogle Scholar
  67. 67.
    Aisen P, Brown EB (1975) Prog Hematol 9:25–56PubMedGoogle Scholar
  68. 68.
    Folajtar DA, Chasteen ND (1982) J Am Chem Soc 104:5775–5780Google Scholar
  69. 69.
    Bell JD, Brown JD, Kubal G, Sadler PJ (1988) Bio Chem Soc Trans 16:714–715Google Scholar
  70. 70.
    Grady JK, Mason AB, Woodworth RC, Chasteen ND (1995) Biochem J 309:403–410PubMedCentralPubMedGoogle Scholar
  71. 71.
    Harris WR, Cafferty AM, Abdollahi S, Trankler K (1998) Biochim Biophys Acta 1383:197–210PubMedGoogle Scholar
  72. 72.
    He QY, Mason AB, Nguyen V, MacGillivray RT, Woodworth RC (2000) Biochem J 350:909–915PubMedCentralPubMedGoogle Scholar
  73. 73.
    Wishnia A, Weber I, Warner RC (1961) J Am Chem Soc 83:2071–2080Google Scholar
  74. 74.
    Byrne SL, Steere AN, Chasteen ND, Mason AB (2010) Biochemistry 49:4200–4207PubMedCentralPubMedGoogle Scholar
  75. 75.
    Egan TJ, Ross DC, Purves LR, Adams PA (1992) Inorg Chem 31:1994–1998Google Scholar
  76. 76.
    Marques HM, Watson DL, Egan TJ (1991) Inorg Chem 30:3758–3762Google Scholar
  77. 77.
    Kretchmar SA, Raymond KN (1988) Inorg Chem 27:1436–1441Google Scholar
  78. 78.
    Williams J, Chasteen ND, Moreton K (1982) Biochem J 201:527–532PubMedCentralPubMedGoogle Scholar
  79. 79.
    Harris WR, Bali PK (1988) Inorg Chem 27:2687–2691Google Scholar
  80. 80.
    Baldwin DA, Egan TJ, Marques HM (1990) Biochim Biophys Acta 1038:1–9PubMedGoogle Scholar
  81. 81.
    Egan TJ, Zak O, Aisen P (1993) Biochemistry 32:8162–8167PubMedGoogle Scholar
  82. 82.
    Marques HM, Walton T, Egan TJ (1995) J Inorg Biochem 57:11–21PubMedGoogle Scholar
  83. 83.
    Zak O, Tam B, MacGillivray RT, Aisen P (1997) Biochemistry 36:11036–11043PubMedGoogle Scholar
  84. 84.
    Li Y, Harris WR (1998) Biochim Biophys Acta 1387:89–102PubMedGoogle Scholar
  85. 85.
    Marques HM, Egan TJ, Pattrick G (1990) S Afr J Sci 86:21–24Google Scholar
  86. 86.
    Muralidhara BK, Hirose M (2000) J Biol Chem 275:12463–12469PubMedGoogle Scholar
  87. 87.
    Mizutani K, Muralidhara BK, Yamashita H, Tabata S, Mikami B, Hirose M (2001) J Biol Chem 276:35940–35946PubMedGoogle Scholar
  88. 88.
    Hamilton DH, Turcot I, Stintzi A, Raymond KN (2004) J Biol Inorg Chem 9:936–944PubMedGoogle Scholar
  89. 89.
    Mazurier J, Spik G (1980) Biochim Biophys Acta 629:399–408PubMedGoogle Scholar
  90. 90.
    Harris WR (1986) J Inorg Biochem 27:41–52PubMedGoogle Scholar
  91. 91.
    Kojima N, Bates GW (1979) J Biol Chem 254:8847–8854PubMedGoogle Scholar
  92. 92.
    Nakazato K, Yamamura T, Satake K (1988) J Biochem 103:823–828PubMedGoogle Scholar
  93. 93.
    Bali PK, Harris WR (1990) Arch Biochem Biophys 281:251–256PubMedGoogle Scholar
  94. 94.
    Turcot I, Stintzi A, Xu J, Raymond KN (2000) J Biol Inorg Chem 5:634–641PubMedGoogle Scholar
  95. 95.
    Hissen AHT, Moore MM (2005) J Biol Inorg Chem 10:211–220PubMedGoogle Scholar
  96. 96.
    Baldwin DA, de Sousa DM (1981) Biochem Biophys Res Commun 99:1101–1107PubMedGoogle Scholar
  97. 97.
    Lehrer SS (1969) J Biol Chem 244:3613–3617PubMedGoogle Scholar
  98. 98.
    Bali PK, Harris WR (1989) J Am Chem Soc 111:4457–4461Google Scholar
  99. 99.
    Halbrooks PJ, Mason AB, Adams TE, Briggs SK, Everse SJ (2004) J Mol Biol 339:217–226PubMedGoogle Scholar
  100. 100.
    Zhang M, Gumerov DR, Kaltashov IA, Mason AB (2004) J Am Soc Mass Spectrom 15:1658–1664PubMedGoogle Scholar
  101. 101.
    Santoro MM, Bolen DW (1988) Biochemistry 27:8063–8068PubMedGoogle Scholar
  102. 102.
    Tanford C (1968) Adv Protein Chem 23:121–282PubMedGoogle Scholar
  103. 103.
    Curtis RA, Lue L (2006) Chem Eng Sci 61:907–923Google Scholar
  104. 104.
    Ramos CHI, Baldwin RL (2002) Protein Sci 11:1771–1778PubMedCentralPubMedGoogle Scholar
  105. 105.
    Zhang Y, Cremer PS (2006) Curr Opin Chem Biol 10:658–663PubMedGoogle Scholar
  106. 106.
    Timasheff SN (1993) Annu Rev Biophys Biomol Struct 22:67–97PubMedGoogle Scholar
  107. 107.
    Nishimura C, Uversky VN, Fink AL (2001) Biochemistry 40:2113–2128PubMedGoogle Scholar
  108. 108.
    van Asselt EJ, Dijkstra BW (1999) FEBS Lett 458:429–435PubMedGoogle Scholar
  109. 109.
    Harris WR (2012) Biochim Biophys Acta 1820:348–361PubMedGoogle Scholar

Copyright information

© SBIC 2014

Authors and Affiliations

  • Sandeep Kumar
    • 1
  • Deepak Sharma
    • 2
  • Rajesh Kumar
    • 1
  • Rajesh Kumar
    • 1
  1. 1.School of Chemistry and BiochemistryThapar UniversityPatialaIndia
  2. 2.Institute of Microbial Technology, Council of Scientific and Industrial ResearchChandigarhIndia

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