Abstract
Elucidating the structure and biosynthesis of neuromelanin (NM) would be an important step towards understanding its putative role in the pathogenesis of Parkinson’s disease. A useful complement to studies aimed at unraveling the origin and properties of this essentially insoluble natural substance is the preparation of synthetic derivatives that resemble NM. With this aim in mind, water-soluble conjugates between dopamine-derived melanin and bovine serum albumin (BSA) were synthesized. Melanin–BSA adducts were prepared with both eumelanic oligomers obtained through the oxidative polymerization of dopamine and pheomelanic oligomers obtained under the same conditions from dopamine and cysteine. Iron ions were added during the synthesis to understand the interaction between the pigment and this metal ion, as the NM in neurons in several human brain regions contains significant amounts of iron. The structures of the conjugates were analyzed by 1H NMR spectroscopy and controlled proteolysis/MS experiments. The binding of iron(III) ions was evaluated by ICP analysis and EPR spectroscopy. The EPR signal from bound iron(III) indicated high-spin octahedral sites and, as also seen for NM, the signal is coupled to a signal from a radical associated with the melanic components of the conjugates. However, the intensity of the EPR signal from iron suggested a reduced fraction of the total iron, indicating that most of the iron is strongly coupled in clusters within the matrix. The amount of paramagnetic, mononuclear iron(III) was greater in the pheomelanin–BSA conjugates, suggesting that iron clustering is reduced in the sulfur-containing pigment. Thus, the melanin–BSA conjugates appear to be good models for the natural pigment.
Similar content being viewed by others
Abbreviations
- BSA:
-
Bovine serum albumin
- DA:
-
Dopamine
- DAQ:
-
Dopaminoquinone
- EDTA:
-
Ethylenediamine tetraacetic acid
- MPTP:
-
1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine
- NM:
-
Neuromelanin
- PD:
-
Parkinson’s disease
References
Hirsch E, Graybiel AM, Agid YA (1988) Nature 334:345–348
Zecca L, Youdim MB, Riederer P, Connor JR, Crichton RR (2004) Nat Rev Neurosci 5:863–873
Fedorow H, Tribl F, Halliday GM, Gerlach M, Riederer P, Double KL (2005) Prog Neurobiol 75:109–124
Zecca L, Costi P, Mecacci C, Ito S, Terreni M, Sonnino S (2000) J Neurochem 74:1758–1765
Ito S, Wakamatsu K (2008) Photochem Photobiol 84:582–592
Zecca L, Shima T, Stroppolo A, Goj C, Battiston GA, Gerbasi R, Sarna T (1996) Swartz HM Neurosci 73:407–415
Shima T, Sarna T, Swartz HM, Stroppolo A, Gerasi R, Zecca L (1997) Free Rad Biol Med 23:110–119
Zecca L, Tampellini D, Gatti A, Crippa R, Eisner M, Sulzer D, Ito S, Fariello R, Gallorini M (2002) J Neural Transm 109:663–672
Aime S, Bergamasco B, Biglino D, Digilio G, Fasano M, Giamello E, Lopiano L (1997) Biochim Biophys Acta 1361:49–58
Zecca L, Gallorini M, Schunemann V, Trautwein AX, Gerlach M, Riederer P, Vezzoni P, Tampellini D (2001) J Neurochem 76:1766–1773
Double KL, Gerlach M, Schunemann V, Trautwein AX, Zecca L, Gallorini M, Youdim BH, Riederer P, Ben-Shachar D (2003) Biochem Pharmacol 66:489–494
Fasano M, Bergamasco B, Lopiano L (2006) J Neural Transm 113:769–774
Zecca L, Casella L, Albertini A, Bellei C, Zucca F, Engelen M, Zadlo A, Szewczyk G, Zareba M, Sarna T (2008) J Neurochem 106:1866–1875
Kropf AJ, Bunker BA, Eisner M, Moss SC, Zecca L, Stroppolo A, Crippa PR (1998) Biophys J 75:3135–3142
Dexter DT, Wells FR, Lees AJ, Agid F, Agid Y, Jenner P, Marsden CD (1989) J Neurochem 52:1830–1836
Jellinger K, Kienzl E, Rumpelmair G, Riederer P, Stachelberger H, Ben-Shachar D, Youdim MB (1992) J Neurochem 59:1168–1171
Faucheux BA, Martin ME, Beaumont C, Hauw JJ, Agid Y, Hirsch EC (2003) J Neurochem 86:1142–1148
Rao KS, Hegde ML, Anitha S, Musicco M, Zucca FA, Turro NJ, Zecca L (2006) Prog Neurobiol 78:364–373
Lindquist NG, Larsson BS, Lyden-Sokolowski A (1998) Neurosci Lett 93:1–6
D’Amato RJ, Lipman ZP, Snyder SH (1986) Science 231:987–989
McGeer PL, Itagaki S, Akiyama H, McGeer EG (1988) Ann Neurol 24:574–576
Langston JW, Forno LS, Tetrud J, Reeves AG, Kaplan JA, Karluk D (1999) Ann Neurol 46:598–605
Zecca L, Wilms H, Geick S, Claasen JH, Brandenburg LO, Holzknecht C, Panizza ML, Zucca FA, Deuschl G, Sievers J, Lucius R (2008) Acta Neuropath 116:47–55
Zhang W, Phillips K, Wielgus AR, Liu J, Albertini A, Zucca FA, Faust R, Qian SY, Miller DS, Chignell CF, Wilson B, Jackson-Lewis V, Przedborski S, Joset D, Loike J, Hong JS, Sulzer D, Zecca L (2011) Neurotox Res 19:63–72
Sulzer D (2007) Trends Neurosci 30:244–250
LaVoie MJ, Ostaszewski BL, Weihofen A, Schlossmacher MG, Selkoe DG (2005) Nat Med 11:1214–1221
Conway KA, Rochet JC, Bieganski RM, Lansbury PT Jr (2001) Science 294:1346–1349
Bridelli MG, Zecca L, Tampellini D (1999) FEBS Lett 457:18–22
d’Ischia M, Napolitano A, Pezzella A, Land EJ, Ramsden CA, Riley PA (2005) Adv Heterocycl Chem 89:1–63
Ito S (1986) Biochim Biophys Acta 883:155–161
Aime S, Bergamasco B, Casu M, Digilio G, Fasano M, Giraudo S, Lopiano L (2000) Mov Disord 15:977–981
Hattori N, Sato S (2007) Neuropathology 27:479–483
Bou-Abdallah F, Chasteen ND (2008) J Biol Inorg Chem 13:15–24
d’Ischia M, Crescenzi O, Pezzella A, Arzillo M, Panzella L, Napolitano A, Barone V (2008) Photochem Photobiol 84:600–607
Meredith P, Sarna T (2006) Pigment Cell Res 19:572–594
Tse DCS, McCreery RL, Adams RN (1976) J Med Chem 19:37–40
Whitehead RE, Ferrer JV, Javitch JA, Justice JB (2001) J Neurochem 76:1242–1251
Nicolis S, Zucchelli M, Monzani E, Casella L (2008) Chem Eur J 14:8861–8873
Peters T Jr (1996) All about albumin: biochemistry genetics and medical applications. Academic, San Diego
Napolitano A, De Lucia M, Panzella L, d’Ischia M (2008) Photochem Photobiol 84:593–599
Gerlach M, Trautwein AX, Zecca L, Youdim MB, Riederer P (1995) J Neurochem 65:923–926
Froncisz W, Sarna T, Hyde JS (1980) Arch Biochem Biophys 202:289–303
Sulzer D, Bogulavsky J, Larsen KE, Behr G, Karatekin E, Kleinman MH, Turro N, Krantz D, Edwards RH, Greene LA, Zecca L (2000) Proc Natl Acad Sci USA 97:11869–11874
Zajac GW, Gallas JM, Cheng J, Eisner M, Moss SC, Alvarado-Swaisgood AE (1994) Biochim Biophys Acta 1199:271–278
Zecca L, Bellei C, Costi P, Albertini A, Monzani Casella L, Gallorini M, Bergamaschi L, Moscatelli A, Turro NJ, Eisner M, Crippa PR, Ito S, Wakamatsu K, Bush WD, Ward WC, Simon JD, Zucca FA (2008) Proc Natl Acad Sci USA 105:17567–17572
Fasano M, Curry S, Terreno E, Galliano M, Fanali G, Narciso P, Notari S, Ascenzi P (2005) IUBMB Life 57:787–796
Di Donato P, Napolitano A (2003) Pigment Cell Res 16:532–539
Simon JD, Peles DN (2010) Acc Chem Res 43:1452–1460
Sulzer D, Mosharov E, Talloczy Z, Zucca FA, Simon JD, Zecca L (2008) J Neurochem 106:24–36
Acknowledgments
The authors thank the Italian MIUR for financial support through a PRIN project, and CIRCMSB.
Author information
Authors and Affiliations
Corresponding author
Electronic supplementary material
Below is the link to the electronic supplementary material.
775_2012_951_MOESM1_ESM.pdf
Proton NMR spectrum of the protein amino acids resulting from complete hydrolysis of a representative melanin–BSA conjugate (Fig. S1); MS/MS spectrum of a dopaminated Cys34 fragment (Fig. S2); EPR spectra of PheoBSA samples (Fig. S3); coverage of the BSA sequence upon proteolytic digestion (Table S1); list of the BSA fragments that were not identified after proteolytic digestion of melanin–BSA conjugates (Table S2). This material is available free of charge via the internet at http://pubs.acs.org (PDF 170 kb)
Rights and permissions
About this article
Cite this article
Ferrari, E., Engelen, M., Monzani, E. et al. Synthesis and structural characterization of soluble neuromelanin analogs provides important clues to its biosynthesis. J Biol Inorg Chem 18, 81–93 (2013). https://doi.org/10.1007/s00775-012-0951-7
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00775-012-0951-7