Hatasa Yukinori, Chikazawa Miho, Furuhashi Mai, Nakashima Fumie, Shibata Takahiro, Kondo Tatsuhiko, Akagawa Mitsugu, Hamagami Hiroki, Tanaka Hiroshi, Tachibana Hirofumi, Uchida Koji
Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya, Japan.
PRESTO, Japan Science and Technology Agency, Saitama, Japan.
PLoS One. 2016 Apr 5;11(4):e0153002. doi: 10.1371/journal.pone.0153002. eCollection 2016.
(-)-Epigallocatechin-3-O-gallate (EGCG), the most abundant polyphenol in green tea, mediates the oxidative modification of proteins, generating protein carbonyls. However, the underlying molecular mechanism remains unclear. Here we analyzed the EGCG-derived intermediates generated upon incubation with the human serum albumin (HSA) and established that EGCG selectively oxidized the lysine residues via its oxidative deamination activity. In addition, we characterized the EGCG-oxidized proteins and discovered that the EGCG could be an endogenous source of the electrically-transformed proteins that could be recognized by the natural antibodies. When HSA was incubated with EGCG in the phosphate-buffered saline (pH 7.4) at 37°C, the protein carbonylation was associated with the formation of EGCG-derived products, such as the protein-bound EGCG, oxidized EGCG, and aminated EGCG. The aminated EGCG was also detected in the sera from the mice treated with EGCG in vivo. EGCG selectively oxidized lysine residues at the EGCG-binding domains in HSA to generate an oxidatively deaminated product, aminoadipic semialdehyde. In addition, EGCG treatment results in the increased negative charge of the protein due to the oxidative deamination of the lysine residues. More strikingly, the formation of protein carbonyls by EGCG markedly increased its cross-reactivity with the natural IgM antibodies. These findings suggest that many of the beneficial effects of EGCG may be partly attributed to its oxidative deamination activity, generating the oxidized proteins as a target of natural antibodies.
(-)-表没食子儿茶素-3-O-没食子酸酯(EGCG)是绿茶中含量最丰富的多酚,它介导蛋白质的氧化修饰,生成蛋白质羰基。然而,其潜在的分子机制仍不清楚。在此,我们分析了与人类血清白蛋白(HSA)孵育时产生的EGCG衍生中间体,并确定EGCG通过其氧化脱氨活性选择性地氧化赖氨酸残基。此外,我们对EGCG氧化的蛋白质进行了表征,发现EGCG可能是天然抗体可识别的电转化蛋白质的内源性来源。当HSA在37°C的磷酸盐缓冲盐水(pH 7.4)中与EGCG孵育时,蛋白质羰基化与EGCG衍生产物的形成有关,如蛋白质结合的EGCG、氧化的EGCG和胺化的EGCG。在体内用EGCG处理的小鼠血清中也检测到了胺化的EGCG。EGCG选择性地氧化HSA中EGCG结合域的赖氨酸残基,生成氧化脱氨产物氨基己二酸半醛。此外,由于赖氨酸残基的氧化脱氨,EGCG处理导致蛋白质负电荷增加。更引人注目的是,EGCG形成的蛋白质羰基显著增加了其与天然IgM抗体的交叉反应性。这些发现表明,EGCG的许多有益作用可能部分归因于其氧化脱氨活性,产生氧化蛋白质作为天然抗体的靶标。