【摘要】 对同步辐射X射线荧光分析技术 (SRXRF)原位测定蛋白质经十二烷基硫酸钠 聚丙烯酰胺凝胶电泳(SDS PAGE)分离后条带中微量元素的方法做了重要改进。通过增加一个干胶步骤 ,使得本底材料对光源的康普顿散射明显减弱 ,本底信号降低到约为湿胶的 1 0 % ,从而可以用来测定蛋白条带内比锌轻的微量元素的相对含量。所提出的方法有望成为一种具有广泛应用前景的生物样品中微量元素的种态分析方法。对人肝细胞胞质溶胶内金属蛋白分布的研究结果表明 :在 1 5～ 95kDa的范围内有 6个含锌蛋白 ,相对分子量分别为1 7.5、2 0 .5、2 7、3 5、5 5及 63kDa ,其中 63kDa蛋白为主要的含锌蛋白 ;此外至少还有 4种分子量分别为 2 0、2 3、4 3和 83 .5kDa的含铁蛋白分布于这个分子量范围内更多还原

【Abstract】 A method for the analysis of trace elements in protein bands with synchrotron radiation X-ray fluorescence (SRXFR) after electrophoresis separation was improved. Through a step of drying the gel before determination, the continues background mainly originated from the Compton scattering of X-ray upon the dry gel matrix was reduced to about one tenth of that upon the wet gel. Trace elements lighter than zinc in protein bands can be determined simultaneously. The method could be used for speciation of trace elements in biological sample. Applying the improved procedure to investigate the distribution of metalloproteins in human liver cytosol, six Zn-containing proteins with molecular weights(MWs) of 17.5, 20.5, 27, 35, 55, and 63 kdalton respectively,and four Fe-containing proteins with MWs of 20, 23, 43, and 83.5 kdalton respectively can be detected. The 63 kdalton Zn-containing protein is the dominant form of zinc in the sample.更多还原