Transglutaminase activity and Nϵ(γ‐glutamyl) lysine isopeptide levels during cell growth: An enzymic and immunological study
Transglutaminase activity and Nϵ(γ‐glutamyl) lysine isopeptide levels during cell growth: An enzymic and immunological study
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细胞生长过程中转谷氨酰胺酶活性和 Nϵ(γ-谷氨酰) 赖氨酸异肽水平：酶学和免疫学研究
DOI：
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发表时间：
1991
影响因子：
6.4
通讯作者：
G. Quash
中科院分区：
文献类型：
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作者：
S. E. Alaoui;S. Legastelois;A. Roch;J. Chantepie;G. Quash
制备了单克隆抗体 (MAb) 81D1c2，该抗体可识别由转谷氨酰胺酶活性作用产生的 Nϵ(γ-谷氨酰) 赖氨酸异肽。它对同源异肽的反应性比 Nα（α 谷氨酰）赖氨酸（一种天然存在的异源二肽）或 Nϵ（γ 谷氨酰）赖氨酸（另一种迄今为止在天然存在的蛋白质中尚未描述的异源肽）的反应性高约 3 倍。当用于对来自人类喉癌 (HEp2) 和鸡胚细胞 (CEC)（均固定在丙酮中）的培养物中的细胞进行免疫组织化学研究时，该 MAb 检测到细胞核中的 Nϵ(γ 谷氨酰) 赖氨酸残基。在 CEC 和 HEp2 细胞生长的滞后期，Nϵ(γ 谷氨酰) 赖氨酸异肽的量密切关注转谷氨酰胺酶活性。然而，在指数生长过程中，HEp2 细胞中的 2 个参数同时降低，而在 CEC 中，转谷氨酰胺酶活性增加，但异肽键水平下降。与其他报道的测量异肽的方法相比，这种免疫组织学方法允许原位定位和至少半定量测定细胞中的 Nϵ(γ 谷氨酰)赖氨酸。
A monoclonal antibody (MAb) 81D1c2, which recognizes the Nϵ(γ‐glutamyl) lysine isopeptide produced by the action of transglutaminase activity was prepared. Its reactivity towards the homologous isopeptide was about 3‐fold greater than that with either Nα (α glutamyl) lysine (a naturally occuring heterologous dipeptide) or Nϵ(γ glutamyl) lysine, another heterologous peptide not described so far in naturally occuring proteins. When used in an immunohistochemical study on cells in culture derived from human carcinoma of the larynx (HEp2) and from chicken embryo cells (CEC), both fixed in acetone, this MAb detected Nϵ(γ glutamyl) lysine residues In the nucleus. The amount of Nϵ(γ glutamyl) lysine isopeptides follows closely transglutaminase activity during the lag phase of growth of both CEC and HEp2 cells. However, during exponential growth, the 2 parameters decrease concomitantly in HEp2 cells, whereas in CEC, transglutaminase activity increases but isopeptide bond levels drop. Compared with other reported methods for measuring isopeptides, this immunohistological approach permits the localization and at least the semi‐quantitative determination of Nϵ(γ glutamyl) lysine in cells in situ.