Nuclear localization of peptidylarginine deiminase V and histone deimination in granulocytes
Nuclear localization of peptidylarginine deiminase V and histone deimination in granulocytes
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DOI：
10.1074/jbc.m208795200
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发表时间：
2002-12-20
影响因子：
4.8
通讯作者：
Yamada, M
中科院分区：
文献类型：
--
作者：
Nakashima, K;Hagiwara, T;Yamada, M
肽精氨酸脱亚胺酶（PAD）可将蛋白中的2个精氨酸残基脱亚胺为瓜氨酸残基。人类有四种pad: I、II、III和V。我们研究了PAD V在HL-60粒细胞和外周血粒细胞中的亚细胞分布。绿色荧光蛋白标记的PAD在HeLa细胞中的表达表明，PAD V定位于细胞核，而PAD I、II和III定位于细胞质。PAD V缺失突变表明序列残基45-74具有核定位信号（NLS）。该NLS的一个序列特征是三个赖氨酸残基簇先于一个脯氨酸残基，这在其他三个pad中没有发现。赖氨酸簇被丙氨酸簇取代，核进口活性被取消。这些结果表明，NLS是典型的单侧NLS。用PAD V特异性抗体染色的HL-60粒细胞、中性粒细胞和嗜酸性粒细胞在细胞核中表现出明显的阳性信号。HL-60粒细胞的亚细胞分离也显示酶的核定位。当中性粒细胞受到钙离子载体A23187的刺激时，细胞核内发生了蛋白质的降解。主要去甲基化蛋白为组蛋白H2A、H3和H4。讨论了PAD V在组蛋白修饰中的意义。
Peptidylarginine deiminase (PAD) deiminates 2 arginine residues in proteins to citrulline residues Ca2+ dependently. There are four types of PADs, I, II, III, and V, in humans. We studied the subcellular distribution of PAD V in HL-60 granulocytes and peripheral blood granulocytes. Expression of green fluorescent protein-tagged PADs in HeLa cells revealed that PAD V is localized in the nucleus, whereas PAD I, II, and III are localized in the cytoplasm. PAD V deletion mutants indicated that the sequence residues 45-74 have a nuclear localization signal (NLS). A sequence feature of this NLS is a three-lysine residue cluster preceded by a proline residue and is not found in the three other PADs. Substitution of the lysine cluster by an alanine cluster abrogated the nuclear import activity. These results suggested that the NLS is a classical monopartite NLS. HL-60 granulocytes, neutrophils, and eosinophils stained with antibody specific for PAD V exhibited distinct positive signals in the nucleus. Subcellular fractionation of HL-60 granulocytes also showed the nuclear localization of the enzyme. When neutrophils were stimulated with calcium ionophore A23187, protein deimination occurred in the nucleus. The major deiminated proteins were identified as histones H2A, H3, and H4. The implication of PAD V in histone modifications is discussed.