Abstract
Surface-enhanced Raman scattering was used as a spectroscopic tool to investigate the changes brought upon cytochrome P450BSß after fatty acid binding. Differences in the spectra of substrate-free and substrate-bound enzyme were observed indicating the potential for this method to be used in the screening of P450 substrates. In particular, the binding characteristics of myristic acid, an inherent substrate, and hydroxylauric acid, a product of fatty acid oxidation, towards P450BSß in the presence of H2O2 were investigated. Specific spectral changes could be assigned to changes in the heme environment only for myristic acid, indicating an occurrence of oxidation process characteristic for the enzymatic substrate.
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Moskovits M (1985) Rev Mod Phys 57:783
Xu H, Aizpurua J, Kall M, Apell P (2000) Phys Rev E 62:4318
Itoh T, Kikkawa Y, Biju V, Ishikawa M, Ikehata A, Ozaki Y (2006) J Phys Chem B 110:2153
Habuchi S, Cotlet M, Gronheid R, Dirix G, Michiels J, Vanderleyden J, De Schryver FC, Hofkens J (2003) J Am Chem Soc 125:8446
Dieringer JA, McFarland AD, Shah NC et al (2006) Faraday Discuss 132:9
Graham D, Faulds F, Smith WE (2006) Chem Commun (Camb) 4363
Hering K, Cialla D, Ackermann K et al (2008) Anal Bioanal Chem 390:113
Stuart DA, Haes AJ, Yonzon CR et al (2005) IEE Proc Nanobiotechnol 152:1
Smith WE (2008) Chem Soc Rev 37:955
Hering KK, Moller R, Fritzsche W et al (2008) Chemphyschem 9:867
Fruk L, Grondin A, Smith WE, et al. (2002) Chem Commun (Camb) 2100
Faulds K, Fruk L, Robson DC et al (2006) Faraday Discuss 132:261
Chourpa I, Lei FH, Dubois P et al (2008) Chem Soc Rev 37:993
Sha MY, Xu H, Penn SG et al (2007) Nanomed 2:725
Aroca RF, Alvarez-Puebla RA, Pieczonka N et al (2005) Adv Colloid Interface Sci 116:45
Hildebrandt P, Stockburger M (1984) J Phys Chem 88:5935
Lee PC, Meisel D (1982) J Phys Chem 86:3391
Lu L, Randjelovic I, Capek R et al (2005) Chem Mat 17:5731
Vogel E, Kiefer W, Deckert V et al (1998) J Raman Spectrosc 29:693
Yu Q, Guan P, Qin D et al (2008) Nano Letters 8:1923
Stöckle RM, Deckert V, Fokas C et al (2000) Appl Spectrosc 54:1577
Han XX, Kitahama Y, Tanaka Y et al (2008) Anal Chem 80:6567
Ingram A, Byers L, Faulds K et al (2008) J Am Chem Soc 130:11846
Ingram AM, Stirling K, Faulds K et al (2006) Org Biomol Chem 4:2869
Moore BD, Stevenson L, Watt A et al (2004) Nat Biotechnol 22:1133
Dunford HB (1999) Heme peroxidases. Wiley, New York
Meunier B, de Visser SP, Shaik S (2004) Chem Rev 104:3947
Bernhardt R (2006) J Biotechnol 124
Guengerich FP (2002) Nat Rev Drug Discov 1:359
Bjerneld EJ, Murty KV, Prikulis J et al (2002) Chemphyschem 3:116
Feis A, Marzocchi MP, Paoli M et al (1994) Biochemistry 33:4577
Murgida DH, Hildebrandt P (2004) Acc Chem Res 37:854
Macdonald IDG, Munro AW, Smith WE (1998) Biophys J 74:3241
Gandubert VJ, Torres E, Niemeyer CM (2008) J Mater Chem 18:3824
Matsunaga I, Ueda A, Sumimoto T et al (2001) Arch Biochem Biophys 394:45
Matsunaga I, Sumimoto T, Ueda A et al (2000) Lipids 35:365
Haines DC, Tomchick DR, Machius M et al (2001) Biochemistry 40:13456
Lee D-S, Yamada A, Sugimoto H et al (2003) J Biol Chem 278:9761
Meunier B (1992) Chem Rev 92:1411
Iain DGM, Smith WE, Andrew WM (1996) FEBS Lett 396:196
Macdonald IDG, Smith WE (1996) Langmuir 12:706
Acknowledgements
We gratefully acknowledge financial support through the Federal Ministry of Education and Research (BMBF) grant “Markerfreie Zelldiagnostik mit Nanometerauflösung” (0312032A and 0312032B). This work was supported in part by the Zentrum für Angewandte Chemische Genomik (ZACG), a joint research initiative founded by the European Union and the Ministry of Innovation and Research of the state North Rhine-Westphalia. CMN thanks Max Planck Society for financial support of a Max Planck Fellow research group at the Max Planck Institute of Molecular Physiology, Dortmund. LF was supported by Marie Curie International Incoming Fellowship (project 514582).
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Bailo, E., Fruk, L., Niemeyer, C.M. et al. Surface-enhanced Raman scattering as a tool to probe cytochrome P450-catalysed substrate oxidation. Anal Bioanal Chem 394, 1797–1801 (2009). https://doi.org/10.1007/s00216-009-2866-x
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DOI: https://doi.org/10.1007/s00216-009-2866-x