Solid-phase synthesis of a pentavalent GalNAc-containing glycopeptide (Tn antigen) representing the nephropathy-associated IgA hinge region.
Solid-phase synthesis of a pentavalent GalNAc-containing glycopeptide (Tn antigen) representing the nephropathy-associated IgA hinge region.
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DOI：
10.1016/j.carres.2010.07.022
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发表时间：
2010-09-23
影响因子：
3.1
通讯作者：
van Die, Irma
中科院分区：
文献类型：
--
作者：
Bolscher, Jan G. M.;Brevoord, Judith;Nazmi, Kamran;Ju, Tongzhong;Veerman, Enno C. I.;van Wijk, Joanna A. E.;Cummings, Richard D.;van Die, Irma
不完全或异常糖基化导致Tn抗原（GalNAcα1-Ser/Thr）在人糖蛋白上的表达与人类病理状况密切相关，包括肿瘤、某些自身免疫性疾病，如特发性IgA肾病，并可能调节免疫稳态。此外，Tn抗原在某些病原体中高度表达，并在宿主-病原体相互作用中发挥作用。为了使实验方法能够研究Tn抗原与免疫系统的相互作用，并分析感染或疾病中的抗Tn抗体反应，我们生成了一个可用于高通量筛选的表达Tn的资源。考虑到铰链区糖基化不完全的IgA肾病，我们使用这个编码五个潜在糖基化位点的铰链序列作为合成表达糖肽的Tn抗原的理想模板。在肽中包含一个n端生物素，可以与链霉亲和素包被的ELISA板结合，使用Helix pomatia凝集素或抗tn单克隆抗体进行监测。我们还发现生物素化的IgA-Tn肽是利用重组t合酶（β1-3-半乳糖转移酶）进行β1-3-半乳糖基化的功能受体。除了其作为IgA肾病可能的诊断工具的免疫化学功能外，该肽是一种极好的聚糖延伸底物，代表了一种适用于聚糖抗原相关疾病的新模板。
Incomplete or aberrant glycosylation leading to Tn antigen (GalNAcα1-Ser/Thr) expression on human glycoproteins is strongly associated with human pathological conditions, including tumors, certain autoimmune diseases, such as the idiopathic IgA nephropathy, and may modulate immune homeostasis. In addition, the Tn antigen is highly expressed by certain pathogens and plays a role in host–pathogen interactions. To enable experimental approaches to study interactions of the Tn antigen with the immune system and analyse anti-Tn antibody responses in infection or disorders, we generated a Tn-expressing resource that can be used for high-throughput screening. In consideration of IgA nephropathy in which the hinge region is incompletely glycosylated, we used this hinge sequence that encodes five potential glycosylation sites as the ideal template for the synthesis of a Tn antigen expressing glycopeptide. Inclusion of an N-terminal biotin in the peptide enabled binding to streptavidin-coated ELISA plates as monitored using Helix pomatia agglutinin or anti-Tn monoclonal antibody. We also found that the biotinylated IgA-Tn peptide is a functional acceptor for β1-3-galactosylation using recombinant T-synthase (β1-3-galactosyltransferase). Besides its immunochemical functionality as a possible diagnostic tool for IgA nephropathy, the peptide is an excellent substrate for glycan elongation and represents a novel template applicable for glycan–antigen-associated diseases.
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影响因子：
11.2
作者：
Ju, Tongzhong;Lanneau, Grainger S.;Cummings, Richard D.
通讯作者：
Cummings, Richard D.
影响因子：
4.8
作者：
Ju, TZ;Brewer, K;Canfield, WM
通讯作者：
Canfield, WM
影响因子：
3.1
作者：
Liu, M;Barany, G;Live, D
通讯作者：
Live, D
影响因子：
4.8
作者：
Ju, TZ;Cummings, RD;Canfield, WM
通讯作者：
Canfield, WM
影响因子：
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作者：
Hiki, Y;Odani, H;Maeda, K
通讯作者：
Maeda, K