Gray J C
Eur J Biochem. 1980 Nov;112(1):39-46. doi: 10.1111/j.1432-1033.1980.tb04984.x.
Cytochrome f has been purified to homogeneity, as judged by polyacrylamide gel electrophoresis, from the leaves of Nicotiana tabacum, Nicotiana glutinosa and their reciprocal hybrids. The cytochrome was extracted from chloroplast membranes by sonication in 2% Triton X-100 and 4M urea and was subsequently purified by acetone precipitation and chromatography on Ultrogel AcA 22 in the presence of cholate and on Sephadex G-200 in the presence of sodium dodecylsulphate. The purified cytochrome had a molecular weight of 32,700 determined by polyacrylamide gel electrophoresis in the presence of sodium dodecylsulphate. A difference in the primary structure of cytochrome f from N. tabacum and N. glutinosa was detected by ion-exchange chromatography of the products of trypsin hydrolysis. Cytochrome f from N. tabacum contained an additional peptide not present in the cytochrome fom N. glutinosa. This additional peptide was present in the cytochrome from N. tabacum female x N. glutinosa male, but not in the cytochrome from N. glutinosa female x N. tabacum male, indicating a maternal mode of inheritance of the primary structure of cytochrome f. This suggests an extranuclear, probably chloroplast, location for the genetic information for cytochrome f.
通过聚丙烯酰胺凝胶电泳判断，细胞色素f已从烟草、粘毛烟草及其正反交杂种的叶片中纯化至同质。该细胞色素通过在2% Triton X-100和4M尿素中超声处理从叶绿体膜中提取，随后通过丙酮沉淀以及在胆酸盐存在下于Ultrogel AcA 22上进行色谱分离和在十二烷基硫酸钠存在下于Sephadex G-200上进行色谱分离来纯化。在十二烷基硫酸钠存在下通过聚丙烯酰胺凝胶电泳测定，纯化后的细胞色素分子量为32,700。通过对胰蛋白酶水解产物进行离子交换色谱分析，检测到烟草和粘毛烟草的细胞色素f一级结构存在差异。烟草的细胞色素f含有一条粘毛烟草的细胞色素f中不存在的额外肽段。这条额外肽段存在于烟草雌性×粘毛烟草雄性的细胞色素中，但不存在于粘毛烟草雌性×烟草雄性的细胞色素中，这表明细胞色素f一级结构存在母系遗传模式。这表明细胞色素f的遗传信息位于核外，可能在叶绿体中。
