Mining the "glycocode"--exploring the spatial distribution of glycans in gastrointestinal mucin using force spectroscopy.
Mining the "glycocode"--exploring the spatial distribution of glycans in gastrointestinal mucin using force spectroscopy.
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DOI：
10.1096/fj.12-221416
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发表时间：
2013-06
期刊：
影响因子：
--
通讯作者：
Juge N
中科院分区：
文献类型：
--
作者：
Gunning AP;Kirby AR;Fuell C;Pin C;Tailford LE;Juge N
粘蛋白是胃肠道粘液层的主要成分。粘蛋白糖基化对大多数分子间和细胞间相互作用至关重要。然而，由于粘蛋白聚糖结构的高度复杂性和异质性，编码的生物信息在很大程度上保持加密。在这里，我们已经开发了一种基于力谱的方法来识别纯化的猪胃粘蛋白（pPGM）和纯化的猪空肠粘蛋白（pPJM）中的生物可及的糖表位。蓖麻凝集素I（RCA），花生（花生）凝集素（PNA），怀槐凝集素II（MALII），和荆豆凝集素I（UEA）的结合特异性利用力光谱测量，以量化的亲和力和空间分布的同源糖在分子尺度上。在pPGM上测定RCA、PNA和UEA的结合能为4、1.6和26 aJ。通过与游离配体的竞争消除结合，证明了亲和力数据的有效性。最近的结合位点分离的分布估计在200 nm粘蛋白片段中的结合位点的数目对于RCA、PNA和UEA为4，对于MALII为1.8。结合位点分离受到pPGM部分去岩藻糖基化的影响。此外，我们表明这种新方法可以解决胃和空肠粘蛋白之间的差异。Gunning，A. P.，Kirby，A. R.，富尔角，Pin，C.，泰尔福德湖E、Juge，N.挖掘“糖密码”--利用力谱技术探索胃肠道粘蛋白中聚糖的空间分布。
Mucins are the main components of the gastrointestinal mucus layer. Mucin glycosylation is critical to most intermolecular and intercellular interactions. However, due to the highly complex and heterogeneous mucin glycan structures, the encoded biological information remains largely encrypted. Here we have developed a methodology based on force spectroscopy to identify biologically accessible glycoepitopes in purified porcine gastric mucin (pPGM) and purified porcine jejunal mucin (pPJM). The binding specificity of lectins Ricinus communis agglutinin I (RCA), peanut (Arachis hypogaea) agglutinin (PNA), Maackia amurensis lectin II (MALII), and Ulex europaeus agglutinin I (UEA) was utilized in force spectroscopy measurements to quantify the affinity and spatial distribution of their cognate sugars at the molecular scale. Binding energy of 4, 1.6, and 26 aJ was determined on pPGM for RCA, PNA, and UEA. Binding was abolished by competition with free ligands, demonstrating the validity of the affinity data. The distributions of the nearest binding site separations estimated the number of binding sites in a 200-nm mucin segment to be 4 for RCA, PNA, and UEA, and 1.8 for MALII. Binding site separations were affected by partial defucosylation of pPGM. Furthermore, we showed that this new approach can resolve differences between gastric and jejunum mucins.—Gunning, A. P., Kirby, A. R., Fuell, C., Pin, C., Tailford L. E., Juge, N. Mining the “glycocode”—exploring the spatial distribution of glycans in gastrointestinal mucin using force spectroscopy.
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DOI：
10.1007/bf01046790
发表时间：
1992-04-01
期刊：
HISTOCHEMICAL JOURNAL
影响因子：
--
作者：
MARTINEZMENARGUEZ, JA;BALLESTA, J;CASTELLS, MT
通讯作者：
CASTELLS, MT
影响因子：
3.4
作者：
Pierres, A;Benoliel, AM;Bongrand, P
通讯作者：
Bongrand, P
影响因子：
3.1
作者：
Jeffers, Faye;Fuell, Christine;Juge, Nathalie
通讯作者：
Juge, Nathalie
影响因子：
1.6
作者：
HUTTER, JL;BECHHOEFER, J
通讯作者：
BECHHOEFER, J
影响因子：
--
作者：
Ceroni A;Dell A;Haslam SM
通讯作者：
Haslam SM