Strong M J, Sopper M M, Crow J P, Strong W L, Beckman J S
John P. Robarts Research Institute, University of Western Ontario, London, Canada.
Biochem Biophys Res Commun. 1998 Jul 9;248(1):157-64. doi: 10.1006/bbrc.1998.8930.
To determine the extent to which enhanced nitration of the low molecular weight neurofilament subunit protein (NFL) is of pathogenic significance in sporadic ALS, we isolated the neurofilament (NF) from the cervical spinal cord of 15 cases of sporadic ALS and 11 age-matched control cases. Of the three NF subunits, only NFL demonstrated consistent nitrotyrosine immunoreactivity on immunoblots against mouse monoclonal anti-nitrotyrosine antibodies. Regardless of whether the NFL was isolated from the Triton X-100 soluble or insoluble cytoskeletal fractions, the extent of NFL nitration did not differ between ALS and control tissue. Similarly, no differences were observed on either two dimensional isoelectric focusing or NFL peptide maps. These findings suggest that NFL is particularly susceptible to peroxynitrite-mediated nitration in vivo, but reveal no significant qualitative or quantitative modifications in the nitration of NFL isolated from sporadic ALS cervical spinal cord tissue as compared to non-ALS controls.
为了确定低分子量神经丝亚基蛋白（NFL）的硝化增强在散发性肌萎缩侧索硬化症（ALS）中具有何种致病意义，我们从15例散发性ALS患者和11例年龄匹配的对照者的颈脊髓中分离出神经丝（NF）。在三种NF亚基中，只有NFL在针对小鼠单克隆抗硝基酪氨酸抗体的免疫印迹上表现出一致的硝基酪氨酸免疫反应性。无论NFL是从Triton X-100可溶性还是不溶性细胞骨架组分中分离出来的，ALS组织和对照组织中NFL的硝化程度均无差异。同样，在二维等电聚焦或NFL肽图上也未观察到差异。这些发现表明，NFL在体内特别容易受到过氧亚硝酸盐介导的硝化作用，但与非ALS对照相比，从散发性ALS颈脊髓组织中分离出的NFL在硝化方面没有明显的定性或定量修饰。
