Terminal N-acetylgalactosamine-specific leguminous lectin from Wisteria japonica as a probe for human lung squamous cell carcinoma.
Terminal N-acetylgalactosamine-specific leguminous lectin from Wisteria japonica as a probe for human lung squamous cell carcinoma.
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DOI：
10.1371/journal.pone.0083886
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Yamamoto K
中科院分区：
文献类型：
--
作者：
Soga K;Teruya F;Tateno H;Hirabayashi J;Yamamoto K
最近根据叶绿体和核DNA序列将崖豆藤重新分类为紫藤属。由于紫藤种子表达的豆科植物凝集素具有独特的N-乙酰半乳糖胺结合特异性，我们纯化紫藤种子凝集素，采用离子交换和凝胶过滤色谱。聚糖微阵列分析表明，与花紫藤和短枝紫藤凝集素结合末端N-乙酰半乳糖胺和半乳糖残基不同，日本紫藤凝集素（WJA）特异性结合α-和β-连接的末端N-乙酰半乳糖胺，但不结合寡糖和糖蛋白上的半乳糖残基。此外，用100多种2-氨基吡啶标记的和对硝基苯基衍生的寡糖进行的正面亲和层析表明，对紫藤凝集素具有最高亲和力的配体是GalNAcβ1-3GlcNAc和GalNAcβ1-4GlcNAc，Ka值分别为9.5 × 104和1.4 × 105 M-1。此外，当在多种细胞系中评估结合时，紫藤凝集素特异性结合EBC-1和HEK 293细胞，而其他紫藤凝集素同等地结合所有测试的细胞系。紫藤凝集素与EBC-1和HEK 293细胞的结合在N-乙酰半乳糖胺存在下显著降低，但半乳糖、甘露糖或N-乙酰葡糖胺不存在，并且通过这些细胞的β-己糖胺酶消化完全消除。这些结果清楚地表明紫藤凝集素与末端N-乙酰半乳糖胺结合，但不与半乳糖结合。此外，对人鳞状细胞癌组织切片的组织化学分析表明，紫藤凝集素特异性结合分化的癌组织，而不是正常组织。紫藤凝集素的这种新的结合特性有可能成为临床应用的有力工具。
Millettia japonica was recently reclassified into the genus Wisteria japonica based on chloroplast and nuclear DNA sequences. Because the seed of Wisteria floribunda expresses leguminous lectins with unique N-acetylgalactosamine-binding specificity, we purified lectin from Wisteria japonica seeds using ion exchange and gel filtration chromatography. Glycan microarray analysis demonstrated that unlike Wisteria floribunda and Wisteria brachybotrys lectins, which bind to both terminal N-acetylgalactosamine and galactose residues, Wisteria japonica lectin (WJA) specifically bound to both α- and β-linked terminal N-acetylgalactosamine, but not galactose residues on oligosaccharides and glycoproteins. Further, frontal affinity chromatography using more than 100 2-aminopyridine-labeled and p-nitrophenyl-derivatized oligosaccharides demonstrated that the ligands with the highest affinity for Wisteria japonica lectin were GalNAcβ1-3GlcNAc and GalNAcβ1-4GlcNAc, with K a values of 9.5 × 104 and 1.4 × 105 M-1, respectively. In addition, when binding was assessed in a variety of cell lines, Wisteria japonica lectin bound specifically to EBC-1 and HEK293 cells while other Wisteria lectins bound equally to all of the cell lines tested. Wisteria japonica lectin binding to EBC-1 and HEK293 cells was dramatically decreased in the presence of N-acetylgalactosamine, but not galactose, mannose, or N-acetylglucosamine, and was completely abrogated by β-hexosaminidase-digestion of these cells. These results clearly demonstrate that Wisteria japonica lectin binds to terminal N-acetylgalactosamine but not galactose. In addition, histochemical analysis of human squamous cell carcinoma tissue sections demonstrated that Wisteria japonica lectin specifically bound to differentiated cancer tissues but not normal tissue. This novel binding characteristic of Wisteria japonica lectin has the potential to become a powerful tool for clinical applications.
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影响因子：
4.3
作者：
Tateno, Hiroaki;Mori, Atsushi;Hirabayashi, Jun
通讯作者：
Hirabayashi, Jun
影响因子：
5.2
作者：
Huang, John;Liang, Jin-Tung;Huang, Min-Chuan
通讯作者：
Huang, Min-Chuan
影响因子：
3.1
作者：
AHMED, H;CHATTERJEE, BP
通讯作者：
CHATTERJEE, BP
影响因子：
2.9
作者：
SUGII, S;KABAT, EA
通讯作者：
KABAT, EA
影响因子：
2.9
作者：
TOYOSHIMA, S;AKIYAMA, Y;OSAWA, T
通讯作者：
OSAWA, T