Site-specific GlcNAcylation of human erythrocyte proteins: potential biomarker(s) for diabetes.
Site-specific GlcNAcylation of human erythrocyte proteins: potential biomarker(s) for diabetes.
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DOI：
10.2337/db08-0994
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发表时间：
2009-02
期刊：
影响因子：
7.7
通讯作者：
Hart, Gerald W.
中科院分区：
文献类型：
--
作者：
Wang, Zihao;Park, Kyoungsook;Comer, Frank;Hsieh-Wilson, Linda C.;Saudek, Christopher D.;Hart, Gerald W.
目的：o -linked N-acetylglucosamine （O-GlcNAc）在糖尿病组织中表达上调，并在胰岛素抵抗和葡萄糖毒性中发挥作用。在这里，我们研究了人红细胞蛋白的glcn酰化程度，并比较了糖尿病人和正常人红细胞蛋白的glcn酰化位点特异性。研究设计和方法-用化学酶法标记和选择性富集葡萄糖酰化红细胞蛋白或葡萄糖酰化肽，并用质谱法鉴定。富集方法结合固相化学衍生化和同位素标记来检测O-GlcNAc修饰位点，并比较正常和糖尿病红细胞蛋白中特定位点的O-GlcNAc占用水平。结果：在人红细胞中检测到催化O-GlcNAc循环（添加和去除）的酶。鉴定了25种糖酰化红细胞蛋白。比较糖尿病红细胞与正常红细胞的蛋白表达水平。可重复鉴定35个O-GlcNAc位点，并计算其位点特异性O-GlcNAc占用率。结论：葡萄糖酰化在红细胞蛋白的不同位点受到不同的调节，以响应血糖状态。这些数据表明，不仅位点特异性的O-GlcNAc水平反映了个体的血糖状态，而且O-GlcNAc在红细胞蛋白上的位点占用可能最终有助于作为早期检测糖尿病的诊断工具。
OBJECTIVE—O-linked N-acetylglucosamine (O-GlcNAc) is upregulated in diabetic tissues and plays a role in insulin resistance and glucose toxicity. Here, we investigated the extent of GlcNAcylation on human erythrocyte proteins and compared site-specific GlcNAcylation on erythrocyte proteins from diabetic and normal individuals. RESEARCH DESIGN AND METHODS—GlcNAcylated erythrocyte proteins or GlcNAcylated peptides were tagged and selectively enriched by a chemoenzymatic approach and identified by mass spectrometry. The enrichment approach was combined with solid-phase chemical derivatization and isotopic labeling to detect O-GlcNAc modification sites and to compare site-specific O-GlcNAc occupancy levels between normal and diabetic erythrocyte proteins. RESULTS—The enzymes that catalyze the cycling (addition and removal) of O-GlcNAc were detected in human erythrocytes. Twenty-five GlcNAcylated erythrocyte proteins were identified. Protein expression levels were compared between diabetic and normal erythrocytes. Thirty-five O-GlcNAc sites were reproducibly identified, and their site-specific O-GlcNAc occupancy ratios were calculated. CONCLUSIONS—GlcNAcylation is differentially regulated at individual sites on erythrocyte proteins in response to glycemic status. These data suggest not only that site-specific O-GlcNAc levels reflect the glycemic status of an individual but also that O-GlcNAc site occupancy on erythrocyte proteins may be eventually useful as a diagnostic tool for the early detection of diabetes.
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影响因子：
4.8
作者：
Kreppel, LK;Hart, GW
通讯作者：
Hart, GW
DOI：
10.1073/pnas.0806216105
发表时间：
2008-09-16
影响因子：
11.1
作者：
Wang, Zihao;Gucek, Marjan;Hart, Gerald W.
通讯作者：
Hart, Gerald W.
DOI：
10.1073/pnas.152346899
发表时间：
2002-08-06
影响因子：
11.1
作者：
McClain, DA;Lubas, WA;Hanover, JA
通讯作者：
Hanover, JA
影响因子：
7
作者：
Wang, Zihao;Pandey, Akhilesh;Hart, Gerald W.
通讯作者：
Hart, Gerald W.
影响因子：
7
作者：
Wells, L;Vosseller, K;Hart, GW
通讯作者：
Hart, GW