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利用新型抗甲硫氨酸亚砜抗体检测选定蛋白质、细胞提取物和血清中的氧化甲硫氨酸

Detection of oxidized methionine in selected proteins, cellular extracts and blood serums by novel anti-methionine sulfoxide antibodies.

作者信息

Oien Derek B, Canello Tamar, Gabizon Ruth, Gasset Maria, Lundquist Brandi L, Burns Jeff M, Moskovitz Jackob

机构信息

Department of Pharmacology and Toxicology, School of Pharmacy, University of Kansas, Lawrence, KS 66045, USA.

出版信息

Arch Biochem Biophys. 2009 May 1;485(1):35-40. doi: 10.1016/j.abb.2009.01.020.

Abstract

Methionine sulfoxide (MetO) is a common posttranslational modification to proteins occurring in vivo.These modifications are prevalent when reactive oxygen species levels are increased. To enable the detection of MetO in pure and extracted proteins from various sources, we have developed novel antibodies that can recognize MetO-proteins. These antibodies are polyclonal antibodies raised against an oxidized methionine-rich zein protein (MetO-DZS18) that are shown to recognize methionine oxidation in pure proteins and mouse and yeast extracts. Furthermore, mouse serum albumin and immunoglobulin (IgG)were shown to accumulate MetO as function of age especially in serums of methionine sulfoxide reductase A knockout mice. Interestingly, high levels of methionine-oxidized IgG in serums of subjects diagnosed with Alzheimer's disease were detected by western blot analysis using these antibodies. It is suggested that anti-MetO-DZS18 antibodies can be applied in the identification of proteins that undergo methionine oxidation under oxidative stress, aging, or disease state conditions.

摘要

甲硫氨酸亚砜(MetO)是蛋白质在体内常见的一种翻译后修饰。当活性氧水平升高时,这些修饰普遍存在。为了能够检测来自各种来源的纯蛋白质和提取蛋白质中的MetO,我们开发了能够识别MetO修饰蛋白质的新型抗体。这些抗体是针对富含氧化甲硫氨酸的玉米醇溶蛋白(MetO-DZS18)产生的多克隆抗体,已证明其能识别纯蛋白质以及小鼠和酵母提取物中的甲硫氨酸氧化。此外,小鼠血清白蛋白和免疫球蛋白(IgG)被证明会随着年龄增长而积累MetO,尤其是在甲硫氨酸亚砜还原酶A基因敲除小鼠的血清中。有趣的是,使用这些抗体通过蛋白质印迹分析检测到,被诊断患有阿尔茨海默病的受试者血清中存在高水平的甲硫氨酸氧化IgG。研究表明,抗MetO-DZS18抗体可用于鉴定在氧化应激、衰老或疾病状态条件下发生甲硫氨酸氧化的蛋白质。

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