Partial Characterization of Proteins from Mussel Mytilus galloprovincialis as a Biomarker of Contamination S. Gorinstein, 1 S.-T. Jung, 2 S. Moncheva, 3 P. Arancibia-Avila, 4 Y.-S. Park, 5 S.-G. Kang, 6 I. Goshev, 7 S. Trakhtenberg, 8 J. Namiesnik 9 1 Department of Medicinal Chemistry and Natural Products, School of Pharmacy, The Hebrew University-Hadassah Medical School, POB 12065, Jerusalem 91120, Israel 2 Department of Food Engineering, Mokpo National University, Muan Jeonnam 534-729, Korea 3 Institute of Oceanology, Bulgarian Academy of Sciences, 9000 Varna, Bulgaria 4 Department of Basic Sciences, Universidad del Bio-Bio, Chillan, Chile 5 Department of Horticultural Science, Mokpo National University, Muan Jeonnam 534-729, Korea 6 Food Industrial Technology Research Center, Mokpo National University, Muan Jeonnam 534-729, Korea 7 Institute of Organic Chemistry and Center of Phytochemistry, Bulgarian Academy of Sciences, 1040 Sofia, Bulgaria 8 Kaplan University Medical Center, Rehovot, Israel 9 Chemical Faculty, Gdansk University of Technology, Gdansk, 80952, Poland Received: 30 November 2004 /Accepted: 9 April 2005 Abstract. Preservation of a healthy environment is a very important task, especially in the time of the total industrial revolution. Therefore, attempts to find new additional bio- markers of contamination are welcomed. For this aim, the functional and antioxidant properties of mussel Mytilus galloprovincialis proteins and the heavy metals Cd and Pb were explored. Mussels were collected in contaminated and noncontaminated sites in the Varna area of the Bulgarian Black Sea coast. Proteins were extracted from mussel entire soft tissue and analyzed using instrumental (Fourier transform infrared [FT-IR] spectroscopy, fluorescence, atomic absorp- tion spectrophotometry), and biochemical (2,2¢-azinobis 3-ethylbenzothiazoline-6-sulfonate) radical cation [ABTS + ]) methods. It was found that mussel proteins from the contam- inated sites had specific qualitative changes: partial unfolding of the a-helix, slight shift in amide I bands, increased hydro- phobicity, and fluorescent intensity in native and denatured samples. In the same mussel samples, an increased radical- scavenging capacity and increased contents of Cd and Pb in entire soft tissue were registered. Therefore, the above-men- tioned indices could be used as additional biomarkers of sea water contamination. Preservation of a healthy environment including prevention of sea contamination is one of the important goals of mankind, especially in times with negative results of industrial devel- opment. In order to control and fight for a healthy environ- ment, monitoring of sea contamination is very important. The influence of different pollutants on mussels and as result some biomarkers of sea pollution were proposed (El Ghazi et al. 2003; Soazig and Marc 2003; Gorinstein et al. 2003; Mon- cheva et al. 2004; Domouhtsidou et al. 2004; Ciocan and Rotchell 2004). Regoli et al. (2004) and Olsson et al. (2004) have described the reaction of the mussel proteins to different pollutants. The response of mussel proteins and their antioxi- dants to marine contamination was demonstrated (Wilhelm et al. 2001; Lopez et al. 2002). Also, protein synthesis regu- lation as a component of cellular stress was used for this purpose (Kalpaxis et al. 2004). The connection between occurrence and distribution of heavy metals in benthic organisms and sea contamination was described by Andreev et al. (1994). Some researchers dem- onstrated that the concentrations of metallothioneins (MTs) can be used as indicator of the degree of sea pollution (El Ghazi et al. 2003; Domouhtsidou et al. 2004; Santamaria- Fernandez et al. 2004; Rainbow et al. 2004). In our most recent investigations (Gorinstein et al. 2003; Moncheva et al. 2004) we have determined the antioxidant activity of different mussel samples and have concluded that total and free poly- phenols can be used as a biomarker of contamination. As was mentioned, Monirith et al. (2003), Olsson et al. (2004), Kalpaxis et al. (2004), and Rainbow et al. (2004) and many others have widely used changes in mussels as biomarkers of sea contamination. Some authors showed that heavy metals that have been used to incubate healthy specimens of the freshwater mussels contributed to a decrease of the metabolic activity, and thus to mineralization in the exposed animals. The most disturbing pollutants were Pb, Zn, Cr, and Cd, which caused greatly decreased overall components, namely, with respect to protein and glucosamine (Moura etal. 2000; Raspor etal. 2004). The main purpose of our study was to control the changes in the protein functionality by the influence of pol- Correspondence to: S. Gorinstein; email: gorin@cc.huji.ac.il Arch. Environ. Contam. Toxicol. 49, 504–510 (2005) DOI: 10.1007/s00244-004-0238-2