使用Sephadex G-75分子篩層析管分離抽取自西班牙三葉草( Lotus purshianus)的部份純化蛋白質和銅結合。結果發現在兩個主要分離群(peak )中，第∥群蛋白和銅結合。比較倆不同品系之西班牙三葉草－耐銅不耐銅，不管有無經過高濃度銅處理，其蛋白質分離型式並無差異，唯銅系經銅處理之根經銅處理後之第∥群蛋白質結合較高濃度之銅，類似銅結合蛋白質。對分離後之第∣、∥群蛋白進行胺基酸成份分析，耐銅系經銅處理後之第∥群蛋白並未如動物的metallothionein一樣有高含量的半膀氨酸(cysteine)，但是卻有較高的天門冬氨酸(aspartate)。計算第∥群蛋白中銅對蛋白質的比例，在銅處理後之耐銅系根為0.135(w/w)，比不耐銅系之根高出約四分之一。Proteins extracted from the roots and shoots of both Cu tolerant and non-tolerant plants of L. purshianus were subjected to Sephadex G-75 column separation. Two major peaks of proteins were found, and the second peak of each separation was found to be associated with Cu by checking Cu in the eluent. There was no difference in the separation pattern between the tolerant and non-tolerant plants, neither is there any difference between the Cu treated and control plants except for the higher copper contents in the peak Ⅱ fractions. Analysis of amino acids composition for peak-Ⅰ and peak-Ⅱ proteins from Cu treated roots did not show a high cysteine content for the copper binding protein (peak-Ⅱ). Peak- Ⅱ proteins were high in acidic amino acids contents. The aspartate contents of peak-Ⅱ from the tolerant plant was higher than that in non-tolerant plant. Copper to protein ratio for the peak-Ⅱ protein in copper treated tolerant roots was 0.135 (w/w), about 25% higher than that in the non-tolerant roots
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