Wilkes D E, Otto J J
Department of Biological Sciences, Purdue University, West Lafayette, IN 47907-1392, USA.
Gene. 2000 Apr 4;246(1-2):295-301. doi: 10.1016/s0378-1119(00)00097-4.
The actin-binding protein profilin was isolated from Tetrahymena thermophila by affinity chromatography, and the peptide sequence was determined for part of the protein. The cDNA sequence was obtained by using the peptide sequence, reverse transcription-PCR and 5' and 3' RACE. The cDNA coded for a profilin of 16680Da, which made it among the largest known profilins, and it had a predicted isoelectric point of 8.27. The deduced amino acid sequence was divergent from other profilins, having more than 26% identity only with profilin from Tetrahymena pyriformis. The sequence contained insertions that are also present in profilins from Tetrahymena pyriformis and Trypanosoma brucei. There appeared to be only a single profilin gene and one transcript from this gene.
通过亲和层析从嗜热四膜虫中分离出肌动蛋白结合蛋白丝切蛋白,并测定了该蛋白部分的肽序列。利用该肽序列、逆转录聚合酶链反应以及5'和3'端快速扩增cDNA末端技术获得了cDNA序列。该cDNA编码一种16680Da的丝切蛋白,这使其成为已知最大的丝切蛋白之一,其预测的等电点为8.27。推导的氨基酸序列与其他丝切蛋白不同,仅与梨形四膜虫的丝切蛋白具有超过26%的同源性。该序列包含在梨形四膜虫和布氏锥虫的丝切蛋白中也存在的插入序列。似乎只有一个丝切蛋白基因,且该基因只有一个转录本。