EXPRESSION OF SEPARATE ISOFORMS OF HUMAN TAU-PROTEIN - CORRELATION WITH THE TAU-PATTERN IN BRAIN AND EFFECTS ON TUBULIN POLYMERIZATION
EXPRESSION OF SEPARATE ISOFORMS OF HUMAN TAU-PROTEIN - CORRELATION WITH THE TAU-PATTERN IN BRAIN AND EFFECTS ON TUBULIN POLYMERIZATION
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DOI：
10.1002/j.1460-2075.1990.tb07870.x
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发表时间：
1990-12-01
期刊：
影响因子：
11.4
通讯作者：
JAKES, R
中科院分区：
文献类型：
--
作者：
GOEDERT, M;JAKES, R
我们在大肠杆菌中表达了六种先前克隆的人微管相关tau蛋白的亚型，并将其纯化为具有生物活性的同质形式。 它们的长度范围为352至441个氨基酸，并且彼此不同之处在于羧基末端一半中存在三个或四个串联重复序列，以及氨基末端中存在或不存在29或58个氨基酸插入物。 当混合在一起时，它们在SDS-PAGE凝胶上产生一组六条带，表观分子量为48-67 kd，具有特征性的间隔模式。 这些条带中的四条与高氯酸提取和碱性磷酸酶处理后成人大脑皮层中发现的主要tau条带对齐。 它们由具有三个重复且无插入的同种型、具有四个重复且无氨基端插入的同种型以及具有29个氨基酸插入的含有三个和四个重复的同种型组成。 在用碱性磷酸酶处理的胎儿人脑提取物中，两条主要tau带之一与含有三个重复的同种型对齐，没有插入，而第二条主要tau带的分子性质仍有待确定。 重组tau亚型在微摩尔浓度下具有生物活性，如通过其促进微管组装的能力所评估的。 与含有三个重复的同种型相比，含有四个重复的同种型的组装速率快2.5-3.0倍，氨基末端插入没有显著贡献。
We have expressed six previously cloned isoforms of human microtubule-associated tau protein in Escherichia coli and purified them to homogeneity in a biologically active form. They range from 352 to 441 amino acids in length and differ from each other by the presence of three or four tandem repeats in the carboxy-terminal half and by the presence or absence of 29 or 58 amino acid inserts in the amino-terminus. When mixed together they gave a set of six bands on SDS-PAGE gels with apparent molecular weights of 48-67 kd and with a characteristic pattern of spacings. Four of these bands aligned with the major tau bands found in adult human cerebral cortex following perchloric acid extraction and alkaline phosphatase treatment. They consisted of isoforms with three repeats and no insertion, four repeats and no amino-terminal insertions and three- and four-repeat containing isoforms with the 29 amino acid insertion. In fetal human brain extracts treated with alkaline phosphatase one of the two major tau bands aligned with the three-repeat containing isoform with no insertions, whereas the molecular nature of the second major tau band remains to be established. The recombinant tau isoforms were biologically active at micromolar concentrations, as assessed by their ability to promote microtubule assembly. The rates of assembly were 2.5-3.0 times faster for isoforms containing four repeats when compared with three-repeat containing isoforms, with no significant contribution by the amino-terminal insertions.