Purification and characterization of recombinant forms of TCL-1 and MTCP-1 proteins.
Purification and characterization of recombinant forms of TCL-1 and MTCP-1 proteins.
复制标题
TCL-1 和 MTCP-1 蛋白重组形式的纯化和表征。
DOI：
10.1006/prep.1997.0822
复制
发表时间：
1998
影响因子：
1.6
通讯作者：
C. M. Croce
中科院分区：
文献类型：
--
作者：
G. C. Du Bois;Sherry P. Song;Irina Kulikovskaya;L. Virgilio;James Varnum;Markus W. Germann;C. M. Croce
TCL-1基因位于14号染色体上，在人类血液系统恶性肿瘤中起重要作用，编码一个14 kDa的蛋白，其功能尚未确定。TCL-1基因在前B细胞、未成熟胸腺细胞和活化T细胞中低水平表达，但在外周成熟B细胞和正常细胞中不表达。TCL-1蛋白在一级结构上与成熟T细胞增殖基因(MTCP-1)编码的蛋白相似。MTCP-1基因位于X染色体上，已被证明与T细胞增生性疾病中罕见的染色体易位有关。TCL-1和MTCP-1基因似乎是与淋巴增殖和T细胞恶性肿瘤有关的基因家族的成员。我们实验室对TCL-1和MTCP-1蛋白进行了研究，以确定这些相关蛋白的结构和功能。在本报告中，我们使用细菌表达系统获得了纯化的TCL-1和MTCP-1蛋白，包括带有和不带有六个His标签序列的形式。重组蛋白经Ni-NTA树脂层析、反相FPLC、pH 7.9的缓冲体系和聚合物基反相柱纯化。与相应的TCL-1蛋白相比，MTCP-1重组蛋白表现出更大的溶解性，不形成二硫键连接的二聚体或低聚物，并且在较低的异丙醇浓度下洗脱。纯化的重组TCL-1和MTCP-1蛋白经N端序列分析、飞行时间质谱仪和圆二色谱鉴定。初步结果表明，去掉His标签的MTCP-1蛋白适用于核磁共振和X射线结晶学结构测定方法。
The TCL-1 gene which is located on chromosome 14 plays a major role in human hematopoeitic malignancies and encodes a 14-kDa protein whose function has not been determined. The TCL-1 gene is expressed in pre-B cells, in immature thymocytes, and at low levels in activated T cells but not in peripheral mature B cells and in normal cells. The TCL-1 protein is similar in its primary structure to a protein encoded by the mature T cell proliferation gene (MTCP-1). The MTCP-1 gene is located on the X chromosome and has been shown to be involved in rare chromosomal translocations in T cell proliferative diseases. The TCL-1 and MTCP-1 genes appear to be members of a family of genes involved in lymphoid proliferation and T cell malignancies. Our laboratory has undertaken the study of the TCL-1 and MTCP-1 proteins to determine the structure and the function of these related proteins. In the present report, we have produced, using a bacterial expression system, both purified TCL-1 and MTCP-1 proteins in forms with and without a six His tag sequence. The recombinant proteins were purified by chromatography on a Ni-NTA resin followed by reverse-phase FPLC using a buffer system at pH 7.9 and a polymeric-based reverse-phase column. The MTCP-1 recombinant proteins display greater solubility, do not form disulfide linked dimers or oligomers, and elute at a lower isopropanol concentration than the corresponding TCL-1 proteins. The purified recombinant TCL-1 and MTCP-1 proteins have been characterized by N-terminal sequence analysis, time of flight mass spectrometry, and circular dichroism spectroscopy. Initial results have indicated that the MTCP-1 protein with the His tag removed is suitable for both NMR and X-ray crystallographic methods of structure determination.
DOI：
10.1016/s0021-9258(18)61070-1
发表时间：
1987-07
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
P. Matsudaira
通讯作者：
P. Matsudaira
编码人类 T 细胞受体 δ 亚基的基因的克隆揭示了其在 α 亚基基因座内的物理组织及其与 T 细胞恶性肿瘤中染色体易位的关系。
DOI：
10.1073/pnas.85.11.3933
发表时间：
1988
影响因子：
11.1
作者：
Isobe,M;Russo,G;Haluska,FG;Croce,CM
通讯作者：
Croce,CM