Deimination of arginine residues in nucleophosmin/B23 and histones in HL-60 granulocytes
Deimination of arginine residues in nucleophosmin/B23 and histones in HL-60 granulocytes
复制标题
DOI：
10.1006/bbrc.2001.6303
复制
发表时间：
2002-01-25
影响因子：
3.1
通讯作者：
Yamada, M
中科院分区：
文献类型：
--
作者：
Hagiwara, T;Nakashima, K;Yamada, M
肽基金氨酸葡萄糖酶（PADS）将蛋白质中的精氨酸残基转化为瓜氨酸残基Ca2+依赖性。 PAD V最近在粒细胞分化的BL-60细胞中发现。为了找到PAD V的靶标，我们将BL-60粒细胞与钙离子载体A23187一起孵育，并通过免疫细胞化学化学研究了脱节的蛋白质，并使用单特种抗体进行了免疫印迹，以修饰的石灰氨水残留物。与A23187孵育后，在核中发现了免疫细胞化学信号。免疫印迹表明40，18-，17-和14 kDa蛋白优先脱离。通过质谱法将40 kDA蛋白聚焦于二维凝胶电泳上的PI 5.0。用0.4 N H2SO4与组蛋白H3，H2A和H4分别提取的18-，17和14 kDa蛋白在二维凝胶电泳上分别与组蛋白专门的二维凝胶电泳。组蛋白的瓜氨酸含量约为组蛋白分子的10％。我们讨论了这些蛋白质对其核功能的能力性的含义。 （C）2002 Elsevier Science（美国）。
Peptidylarginine deiminases (PADS) convert arginine residues in proteins into citrulline residues Ca2+-dependently. PAD V was recently found in granulocyte-differentiated BL-60 cells. To find a target of PAD V, we incubated BL-60 granulocytes with the calcium ionophore A23187 and studied deiminated proteins by immunocytochemistry and immunoblotting using a monospecific antibody to modified citrulline residues. Immunocytochemical signals were found in the nucleus upon incubation with A23187. Immunoblotting indicated that 40-, 18-, 17-, and 14-kDa proteins were preferentially deiminated. The 40-kDa protein, which was focused to pI 5.0 on two-dimensional gel electrophoresis, was identified as nucleophosmin/B23 by mass spectrometry. The 18-, 17-, and 14-kDa proteins extracted with 0.4 N H2SO4 comigrated with histones H3, H2A, and H4, respectively, on two-dimensional gel electrophoresis specialized for histones. The citrulline content of histones amounted to about 10% of the histone molecules. We discuss the implications of deimination of these proteins for their nuclear functions. (C) 2002 Elsevier Science (USA).