Quantification of fucosylated hemopexin and complement factor H in plasma of patients with liver disease.
Quantification of fucosylated hemopexin and complement factor H in plasma of patients with liver disease.
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DOI：
10.1021/ac502727s
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发表时间：
2014-11-04
影响因子：
7.4
通讯作者：
Goldman, Radoslav
中科院分区：
文献类型：
--
作者：
Benicky, Julius;Sanda, Miloslav;Pompach, Petr;Wu, Jing;Goldman, Radoslav
增强的聚焦已被认为是肝脏疾病和肝细胞癌（HCC）血清学监测的标志。我们提出了一种定量位点特异性分析聚焦化的工作流程，并将其应用于健康个体和肝硬化和HCC患者的两种肝分泌糖蛋白——血凝素（HPX）和补体因子H （CFH）的比较。纯化后的糖蛋白衍生的糖肽在混合样品（2池/组，5个样品/池）上进行无标记LC-MS定量，并分别使用唾液酸酶和内糖苷酶F2/F3进行糖苷酶辅助分析，以提高糖型的分辨率。我们的分析显示，个体聚焦型糖的相对丰度归一化到其非聚焦型糖的水平，揭示了肝脏疾病中聚焦化的持续增加，具有显著的位点和蛋白质特异性差异。我们观察到HPX的N187位点糖型的微观异质性最高，CFH的N882和N911位点没有核心聚焦，或者CFH的核心聚焦程度高于HPX，但我们没有发现HCC与匹配的肝硬化样本之间的差异。糖苷酶辅助LC-MS-MRM分析通过简化方案制备的个体患者样品证实了数量差异。外臂病灶特异性转移记录了HPX N187位点双天线和三天线聚糖外臂病灶的疾病相关增加。HPX和CFH的聚焦增强可能是恶性前肝病的一个指标，需要进一步验证。分析策略可以很容易地适应于分析其他蛋白质在适当的疾病背景。
Enhanced fucosylation has been suggested as a marker for serologic monitoring of liver disease and hepatocellular carcinoma (HCC). We present a workflow for quantitative site-specific analysis of fucosylation and apply it to a comparison of hemopexin (HPX) and complement factor H (CFH), two liver-secreted glycoproteins, in healthy individuals and patients with liver cirrhosis and HCC. Label-free LC-MS quantification of glycopeptides derived from these purified glycoproteins was performed on pooled samples (2 pools/group, 5 samples/pool) and complemented by glycosidase assisted analysis using sialidase and endoglycosidase F2/F3, respectively, to improve resolution of glycoforms. Our analysis, presented as relative abundance of individual fucosylated glycoforms normalized to the level of their nonfucosylated counterparts, revealed a consistent increase in fucosylation in liver disease with significant site- and protein-specific differences. We have observed the highest microheterogeneity of glycoforms at the N187 site of HPX, absence of core fucosylation at N882 and N911 sites of CFH, or a higher degree of core fucosylation in CFH compared to HPX, but we did not identify changes differentiating HCC from matched cirrhosis samples. Glycosidase assisted LC-MS-MRM analysis of individual patient samples prepared by a simplified protocol confirmed the quantitative differences. Transitions specific to outer arm fucose document a disease-associated increase in outer arm fucose on both bi- and triantennary glycans at the N187 site of HPX. Further verification is needed to confirm that enhanced fucosylation of HPX and CFH may serve as an indicator of premalignant liver disease. The analytical strategy can be readily adapted to analysis of other proteins in the appropriate disease context.
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DOI：
10.1158/1078-0432.ccr-07-5261
发表时间：
2009-03-01
期刊：
Clinical cancer research : an official journal of the American Association for Cancer Research
影响因子：
--
作者：
Goldman R;Ressom HW;Varghese RS;Goldman L;Bascug G;Loffredo CA;Abdel-Hamid M;Gouda I;Ezzat S;Kyselova Z;Mechref Y;Novotny MV
通讯作者：
Novotny MV
影响因子：
3.7
作者：
Comunale MA;Rodemich-Betesh L;Hafner J;Wang M;Norton P;Di Bisceglie AM;Block T;Mehta A
通讯作者：
Mehta A
影响因子：
8.8
作者：
Johnson PJ;Poon TC;Hjelm NM;Ho CS;Ho SK;Welby C;Stevenson D;Patel T;Parekh R;Townsend RR
通讯作者：
Townsend RR
影响因子：
13.5
作者：
Liu, Xue-En;Desmyter, Liesbeth;Chen, Cuiying
通讯作者：
Chen, Cuiying
影响因子：
4.4
作者：
Lee, Hyoung-Joo;Cha, Hyun-Jeong;Paik, Young-Ki
通讯作者：
Paik, Young-Ki