Amino acid residues 226-240 of tau, which encompass the first Lys-Ser-Pro site of tau, are partially phosphorylated in Alzheimer paired helical filament-tau.
Amino acid residues 226-240 of tau, which encompass the first Lys-Ser-Pro site of tau, are partially phosphorylated in Alzheimer paired helical filament-tau.
复制标题
tau 的氨基酸残基 226-240（包含 tau 的第一个 Lys-Ser-Pro 位点）在阿尔茨海默氏症配对螺旋丝 tau 中部分磷酸化。
DOI：
10.1046/j.1471-4159.1994.62031055.x
复制
发表时间：
1994
影响因子：
4.7
通讯作者：
Yen,SH
中科院分区：
文献类型：
--
作者：
Liu,WK;Dickson,DW;Yen,SH
使用对应于人τ的残基226-240（E9肽）的合成肽（其含有Lys-Ser-Pro基序）来产生多克隆抗体。抗体E9与磷酸化E9肽的反应性比与天然E9肽的反应性低10倍。E9抗体用于研究在阿尔茨海默氏病脑中发现的τ（PHF-τ）的修饰形式中的磷酸化程度，并掺入成对螺旋丝（PHF）中。E9免疫标记的阿尔茨海默病神经元缠结和异常的神经突起在脑切片强烈，与磷酸酶预处理后的切片检测到的免疫反应性增加。在免疫印迹和ELISA上，E9与PHF-τ和重组人τ反应，但不与高和中分子量神经丝蛋白反应。PHF‐τ的磷酸酶处理使E9免疫反应性提高了30- 50%。去磷酸化的高分子量但不中等分子量的神经丝蛋白与E9反应。这些结果表明，在对应于τ残基226-240的亚区中，<50%的PHF-T被磷酸化，并表明该区域的磷酸化可能不是PHF形成所必需的。
A synthetic peptide corresponding to residues 226–240 (E9 peptide) of human τ, which contains an Lys‐Ser‐Pro motif, was used to raise a polyclonal antibody. The antibody, E9, was 10‐fold less reactive with phospho‐E9 peptide than with native E9 peptide. E9 antibody was used to study the extent of phosphorylation in a modified form of τ (PHF‐τ) that is found in Alzheimer's disease brain and is incorporated into paired helical filaments (PHFs). E9 immunolabeled Alzheimer's disease neurofibrillary tangles and abnormal neurites in brain sections intensely, with increased immunoreactivity detected after pretreatment of sections with phosphatase. On immunoblots and ELISA, E9 reacted with PHF‐τ and recombinant human τ but not with the high and middle molecular weight neurofilament proteins. Phosphatase treatment of PHF‐τ improved the E9 immunoreactivity by 30–50%. Dephosphorylated high but not middle molecular weight neurofilament protein became reactive with E9. These results indicate that <50% of the PHF‐T is phosphorylated in the subregion corresponding to residues 226–240 of τ and suggest that the phosphorylation of this region may not be essential for PHF formation.