ISSN 100727626 CN 1123870ΠQ 2003 6 Chinese Journal of Biochemistry and Molecular Biology 19 (3) :377 382, 3,, (, 918 100039), (, 550002) (Sephadex G2100),, ;, ;, 11 kd (MT),,,,,,,,, Q51,X132 Distribution of Mercury and Selenium in the Proteins of Porcine Liver and Kidney Under Different Mercury Exposure Level ZHAO Jiu2jiang, ZHANG Pei2qun 3,CHEN Chun2ying, CHAI Zhi2fang ( Laboratory of Nuclear Analytical Techniques, Institute of High Energy Physics, Chinese Academy of Sciences, Beijing 100039, China) QU Li2ya, LI Mei ( Guizhou Research and Designing Institute of Environmental Sciences, Guiyang 550002, China) Abstract The mercury and selenium in cytosolic proteins porcine liver and kidney from Guizhou and Beijing were determined by atomic fluorescence spectrometry after separation with Sephadex G2100 gel filtration chromatography The results showed that the distribution patterns of selenium and mercury in the cytosolic proteins from the two area samples were different Selenium was mainly bound to the cytosolic proteins with :2002209220, :2002212210 (No 19935020 No 20071032) (No KJCX22N101) 3 Tel : (010) 88233212,E2mail :zhangpq @ihep ac cn,,1974 7, Received :September 20,2002 ;Accepted :December 10,2002 Supported by the National Natural Science Foundation of China (No 19935020 and No 20071032) and the Major Project of Chinese Academy of Sciences (No KJCX22N101) 3 Corresponding author Tel : (010) 88233212,E2mail :zhangpq @ihep ac cn
378 19 high molecular weight in Beijing porcine liver, while it was also present with low molecular weight fractions besides being bond to the high molecular weight proteins for the Guizhou one Mercury was mainly bound to the proteins with high molecular weight for the Guizhou porcine liver Se2containing components existed in the fractions of both high and low molecular weight for the Guizhou porcine kidney, while they were only present in the fractions of high molecular weight for Beijing one Mercury was mainly in the form of metallothioneins (MTs) in the cytosol of Beijing porcine kidney On the contrary, it was mainly bound to the proteins of higher molecular weight besides to MTs for the Guizhou one The present results indicate that the different level of mercury exposure results in not only the different contents of selenium and mercury in the cytosolic proteins of porcine liver and kidney from the two areas, but also their different distribution patterns, suggesting that there is a certain correlation between the two elements in the metabolism of organisms Key words mercury, selenium, porcine tissues, atomic fluorescence spectrometry,distribution, ( n = 100, m = 35), ( ),, [1 ],, [2 ] 1 1, 1975 Koeman [3 ],,,, [4 ] 1 Imura,,,, 1 1, [7,8 1 min, 5 % ], ;, 50 % 10 ml 011 molπl ( ph 816), 20 mgπml, Sephadex G2100, 15 mlπh, 5 ml, 1 1 [2 ] Falnoga [5 ], 1, 2, AFS2820 [9 280 nm 220 nm ] 2, 3 1, 110 120 5 molπl, Se ( ) Se,Suzuki [6 ] ( ), NaBH 4 P { (Se2Hg) n } m 2 P Se ( ),
3 : 379, 3 1, 110,,,,,,,,, Table 1, Table 1 Determination of Se and Hg in some standard reference materials(srm) with atomic fluorescence spectrometry ( gπg dry weight) SRM Se measured value a Se certified value a Hg measured value a Hg certified value a NIST 1577a GBW 08571 IAEA H28 GBW 080193 0168 0106 0171 0103 3 3128 0115 3165 0117 3 3 01060 01020 01067 01008 3 3 4127 0101 4167 0129 3 3 3 0188 0103 0191 0108 3 3 3 0153 0106 0156 0107 GBW 08572 01212 01003 01201 01004 3 3 GBW 09101 0151 0109 0158 0105 3 3 2105 0107 2116 0121 3 3 a Data represent means and standard deviation of 2 4 independent SRM samples 3 I Roelandts et al, 1998 [10 ], 3 3 IAEA, 1995 [11 ], 3 3 3 IAEA, 1985 [12 ] 2 211 91 15 kd 412 011 kd, Fig11 Fig 2 2, 98 7 kd, 119 011 kd 41 3 kd,311 011 kd, Fig11 Fig 2, 103 kd Fig 1 Distribution of Se, Hg and proteins in the eluate of porcine liver cytosol from Guizhou after separation with Sephadex G2100 gel filtration chromatography Se ; Hg ; A 280
380 19 81 % ; 33 12 kd (0149 gπg), 1317, (1519 gπg), % 513 %,,,,, (01063 gπg), (1110, gπg), Fig 2 Distribution of Se, Hg and proteins in porcine liver cytosol from Beijing after separation with Sephadex G2100 gel filtration chromatography Se ; Hg ; A 280 kd 57 7 kd 212, Fig13 Fig14 105 5 kd,418 015 Fig 3 Distribution of Se, Hg and proteins in the eluate of porcine kidney cytosol from Guizhou after separation with Sephadex G2100 gel filtration chromatography Se ; Hg ; A 280 ; 222222222 A 220
3 : 381 108 kd,75 kd 415 kd, (0124 gπ 3511 %,3010 % 2914 % g), (2112 gπg),101 kd 7112 %, ;, (3169 gπg),, (1918 gπg), [1 ] (, ),, Fig 4 Distribution of Se, Hg and proteins in the eluate of porcine kidney cytosol from Beijing after separation with Sephadex G2100 gel filtration chromatography Se ; Hg ; A 280 ; 222222222 A 220,, 108 kd 11 kd,, 72 % 28 % 11 kd, 3215 %, MTs, 213,,, 11, kd 11 kd 280 nm,, 220 nm, (MTs) (915 1215 kd) MT 11 kd,,, [13 220 nm, 280 nm ],,,,, ; Chen [7 ],
382 19 10 % 211 218, 6 Suzuki K T, Sasakura C, Yoneda S, Binding sites for the, 1998,1429 : 102 112 7 Chen C, Zhang P, Hou X, Chai Z Investigation of selenium distribution in subcellular fractions of human liver by neutron 131 138 8,,, ( References) 1 Zhi2fang Isolation and determination of selenium2containing : ( Xu Hui2bi ed Chemistry and Biochemistry of Selenium and Its Application in Life Sciences Technology Press), 1994 Wuhan : Huazhong University of Science and 2 ( 2 ) : ( Wang Kui ed Trace Elements in Life sciences 2 nd ed Beijing : Chinese Metrology Press), 1996 3 Koeman J H, Van de ven W S M, de Goeij J J M, Tjioe P S, van Haaften J L Mercury and selenium in marine mammals and birds Sci Total Environ, 1975, 3 : 279 287 4 Kosta L, Byrne A, Zelenko V Correlation between selenium and mercury in man following exposure to inorganic mercury Nature, 1975,254 : 238 239 5 Falnoga I, Tusek2Znidaric M, Horvat M, Stegnar P, Mercury, selenium, and cadmium in human autopsy samples from Idrija residents and mercury mine works Environ Res, 2000, 84 : ( Hg2Se) complex on selenoprotein P Biochim Biophys Acta, activation analysis Biol Trace Element Res, 1999, 71272 : ( Zhao Jiu2jiang, Chen Chun2ying, Zhang Pei2qun, Chai proteins in subcellular fractions of human liver Chin J Biochem Mol Biol), 2001, 17 (6) :800 803 9,,,,, : (Li Jian2wu, Xiao Neng2qing, Yu Rui2yuan, Yuan Ming2xiu, Chen Li2rong, Chen Ya2hui, Chen Lai2tong ed Experimental Theories and Methods in Biochemistry Beijing : Peking University Press), 1994 : 82 91 10 Roelandts I, Glandney E S Consensus values for NIST biological and environmental Fresenius J Anal Chem, 1998,360 : 327 338 standard reference materials 11 IAEA2TECDOC2854, survey of reference materials, 1995 12 IAEA, IAEA horse kidney provisional certificate of analysis, 1985 13 Vasak M Criteria of purity for metallothioneins Methods in Enzymology, 1991,205 : 45 47