Lin C H, Wu M
Institute of Molecular Biology, Academia Sinica, Nankang, Taipei, Republic of China.
Plant Mol Biol. 1990 Sep;15(3):449-55. doi: 10.1007/BF00019161.
Previously, a ferredoxin-type iron-sulfur protein, frx B protein, was identified in a high-salt extract of the purified thylakoid membrane of Chlamydomonas reinhardtii, a unicellular green alga. Polyclonal antibody was raised against a synthetic pentadecameric peptide with an amino acid sequence corresponding to the highly conserved region of the putative frx B proteins of 3 land plants. In this report, protein(s) reacting strongly and specifically with this antibody was detected in the equivalent high-salt extract prepared from purified chloroplast of spinach and tobacco. One strong reaction polypeptide band from tobacco chloroplast was purified from SDS-polyacrylamide gel and subjected to endoproteinase lys C digestion. The resulting polypeptides were separated by reversed-phase chromatography. N-terminal sequencing of 3 purified polypeptides revealed that the protein is encoded by the 'frxB gene' identified from DNA sequence analysis.
此前，在单细胞绿藻莱茵衣藻纯化类囊体膜的高盐提取物中鉴定出一种铁氧化还原蛋白型铁硫蛋白，即frx B蛋白。针对一种合成的十五聚体肽制备了多克隆抗体，该肽的氨基酸序列对应于3种陆地植物假定的frx B蛋白的高度保守区域。在本报告中，在从菠菜和烟草的纯化叶绿体制备的等效高盐提取物中检测到了与该抗体发生强烈且特异性反应的蛋白质。从烟草叶绿体的一条强反应性多肽带在SDS-聚丙烯酰胺凝胶上进行纯化，并进行内肽酶赖氨酸C消化。所得多肽通过反相色谱法分离。对3条纯化多肽的N端测序表明，该蛋白由DNA序列分析鉴定的“frxB基因”编码。
