IDENTIFICATION OF THE MAJOR MULTIPHOSPHORYLATION SITE IN MAMMALIAN NEUROFILAMENTS
IDENTIFICATION OF THE MAJOR MULTIPHOSPHORYLATION SITE IN MAMMALIAN NEUROFILAMENTS
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DOI：
10.1073/pnas.85.6.1998
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发表时间：
1988-03-01
影响因子：
11.1
通讯作者：
LAZZARINI, RA
中科院分区：
文献类型：
--
作者：
LEE, VMY;OTVOS, L;LAZZARINI, RA
序列 Lys-Ser-Pro-Val-Pro-Lys-Ser-Pro-Val-Glu-Glu-Lys-Gly 在人类中型神经丝 (NF) 蛋白 (NF-M) 中连续重复六次。为了确定 Lys-Ser-Pro-Val(Ala) 是否是体内 NF 磷酸化的主要位点，合成了基于人 NF-M 重复序列的肽并进行了化学磷酸化。这些合成肽用 515 种单克隆抗体 (mAb) 进行探测，这些单克隆抗体针对多种不同磷酸化形式的 NF 蛋白进行了检测和区分。对其中 95 种在化学磷酸化前后识别肽的 mAb 进行的研究表明，这些肽共有的高度免疫原性表位存在于所有测试物种（包括无脊椎动物）的 NF 中。这表明主要 NF 磷酸化位点的系统发育保守性。最后，这些肽和主要 NF 磷酸化位点共有的交叉反应抗原决定簇被证明存在于阿尔茨海默病患者的神经原纤维缠结中，以及两种神经元特异性微管相关蛋白 (MAP)（即 MAP2 和 tau）中。
The sequence Lys-Ser-Pro-Val-Pro-Lys-Ser-Pro-Val-Glu-Glu-Lys-Gly repeats six times serially in the human midsized neurofilament (NF) protein (NF-M). To establish whether Lys-Ser-Pro-Val(Ala) is the major site for in vivo NF phosphorylation, peptides based on the human NF-M repeat were synthesized and chemically phosphorylated. These synthetic peptides were probed with 515 monoclonal antibodies (mAbs) that were raised to, and distinguished, several differentially phosphorylated forms of NF proteins. Studies with 95 of those mAbs that recognized the peptides before and after chemical phosphorylation demonstrated that a highly immunogenic epitope shared by the peptides is present in NFs from all species tested, including invertebrates. This suggests the phylogenetic conservation of a major NF phosphorylation site. Lastly, a cross-reactive antigenic determinant shared by the peptides and the major NF phosphorylation site was shown to exist in neurofibrillary tangles of patients with Alzheimer disease as well as in two neuron-specific microtubule-associated proteins (MAPs)--i.e., MAP2 and tau.