Actin carbonylation: From a simple marker of protein oxidation to relevant signs of severe functional impairment
Actin carbonylation: From a simple marker of protein oxidation to relevant signs of severe functional impairment
复制标题
DOI：
10.1016/s0891-5849(01)00690-6
复制
发表时间：
2001-11-01
影响因子：
7.4
通讯作者：
Colombo, R
中科院分区：
文献类型：
--
作者：
Dalle-Donne, I;Rossi, R;Colombo, R
蛋白质结合的羰基的数量是蛋白质氧化的既定标志。最近的证据表明，阿尔茨海默病患者的大脑和缺血性心脏中的肌动蛋白羰基含量显着增加。在人类结肠细胞暴露于次氯酸 (HOCI) 后，肌动蛋白羰基化增强，导致肌动蛋白细胞骨架破坏和屏障功能丧失。在此，介绍了 HOCI 诱导的氧化对纯化肌动蛋白的影响。结果表明 HOCI 导致羰基产率快速增加。然而，当羰基化变得明显时，一些半胱氨酸和蛋氨酸残基已经被氧化。已经发现了羰基化肌动蛋白的共价分子间交联以及一些非共价聚集。共价交联不受还原剂和变性剂的影响，与二酪氨酸荧光的增加平行。此外，HOCI介导的氧化诱导肌动蛋白丝的逐渐破坏和F-肌动蛋白形成的抑制。导致肌动蛋白聚合抑制的HOCI与肌动蛋白的摩尔比似乎仅对半胱氨酸和甲硫氨酸有影响。涉及氨基酸侧链氧化并形成羰基的过程将在氧化损伤程度高于导致某些关键氨基酸残基氧化的程度下发生。因此，体内发现的肌动蛋白羰基含量的增加表明剧烈的氧化修饰导致剧烈的功能损伤。 (C) 2001 爱思唯尔科学公司。
The number of protein-bound carbonyl groups is an established marker of protein oxidation. Recent evidence indicates a significant increase in actin carbonyl content in both Alzheimer's disease brains and ischemic hearts. The enhancement of actin carbonylation, causing the disruption of the actin cytoskeleton and the loss of the barrier function, has also been found in human colonic cells after exposure to hypochlorous acid (HOCI). Here, the effects of oxidation induced by HOCI on purified actin are presented. Results show that HOCI causes a rapidly increasing yield of carbonyl groups. However, when carbonylation becomes evident, some Cys and Met residues have been already oxidized. Covalent intermolecular cross-linking as well as some noncovalent aggregation of carbonylated actin have been found. The covalent cross-linking, unaffected by reducing and denaturing agents, parallels an increase in dityrosine fluorescence. Moreover, HOCI-mediated oxidation induces the progressive disruption of actin filaments and the inhibition of F-actin formation. The molar ratios of HOCI to actin that lead to inhibition of actin polymerization seem to have effect only on cysteines and methionines. The process that involves oxidation of amino acid side chains with formation of a carbonyl group would occur at an extent of oxidative insult higher than that causing the oxidation of some critical amino acid residues. Therefore, the increase in actin content of carbonyl groups found in vivo would indicate drastic oxidative modification leading to drastic functional impairments. (C) 2001 Elsevier Science Inc.