Purification and characterization of ribonucleoproteins from pea chloroplasts.
Purification and characterization of ribonucleoproteins from pea chloroplasts.
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豌豆叶绿体核糖核蛋白的纯化和表征。
DOI：
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发表时间：
1993
期刊：
影响因子：
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通讯作者：
K. Tewari
中科院分区：
文献类型：
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作者：
C. Subbaiah;K. Tewari
已知 RNA 结合蛋白可介导许多生物体中基因的转录后调控。最近发现它们在质体基因的表达中很重要。我们使用单链 DNA 亲和层析从豌豆 (Pisum sativum L.) 叶绿体提取物中纯化了一组三种单链核酸特异性酸性蛋白（33、30 和 28 kDa）。它们都具有酸性氨基末端，但每种多肽的氨基酸序列都是独特的，与最近报道的来自烟草叶绿体的核糖核蛋白有部分相似性。使用纯化蛋白的多克隆抗血清进行蛋白质印迹分析表明，豌豆蛋白的抗原性也不同。此外，根据它们的大核酸结合结构域和多核苷酸底物亲和力，预测它们属于豌豆质体核糖核蛋白家族。在存在翻译抑制剂的情况下对蛋白质进行体内放射性标记以及叶组织RNA的体外翻译表明这些蛋白质是在细胞核中编码的。抗体交叉反应实验表明，它们的基因在质体进化过程中是保守的。
RNA-binding proteins are known to mediate the post-transcriptional regulation of genes in many organisms. Recently they have been found to be important in the expression of plastid genes. We have purified a group of three single-stranded nucleic-acid-specific acidic proteins (33, 30 and 28 kDa) from chloroplast extracts of pea (Pisum sativum L.), using single-stranded DNA affinity chromatography. All of them have acidic amino termini but the amino acid sequences are unique to each polypeptide, with partial similarities to the recently reported ribonucleoproteins from tobacco chloroplasts. The pea proteins are also antigenically distinct, as shown by Western blot analysis using polyclonal antisera for purified proteins. Further, from their large nucleic-acid-binding domains and the polynucleotide substrate affinities, they are predicted to belong to a family of pea plastid ribonucleoproteins. In vivo radiolabeling of proteins in the presence of translational inhibitors as well as in vitro translation of leaf tissue RNA suggest that these proteins are encoded in the nucleus. Antibody cross-reactivity experiments reveal that their genes are conserved during plastid evolution.