Single chain antibodies that recognize the N-glycosylation site.
Single chain antibodies that recognize the N-glycosylation site.
复制标题
识别 N-糖基化位点的单链抗体。
DOI：
10.1016/j.abb.2003.12.032
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发表时间：
2004
影响因子：
3.9
通讯作者：
Toshiki Tanaka
中科院分区：
文献类型：
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作者：
M. Kikuchi;Mizuho Kataoka;T. Kojima;T. Horibe;Kan Fujieda;Taiji Kimura;Toshiki Tanaka
我们的目的是鉴定可以识别指导寡糖基转移酶（OT）活性的Asn-Xaa-Ser/Thr（NXS/T）N-糖基化位点的抗体。我们使用合成的Asn-Cys-Ser/Thr（NCS/T）三肽与牛血清白蛋白缀合，从Griffin 1噬菌体抗体库中分离可变区的单链抗体片段（scFv）。虽然Ser和Thr具有不同的侧链，但是这样分离的scFv蛋白质以10− 6 M量级的Kd值与NCS和NCT结合，并且接受Cys残基被各种氨基酸（包括Ala、Gly和瓦尔）取代。然而，这些蛋白既不识别Asn-Pro-Ser/Thr也不识别非NXS/T三肽。当scFv蛋白处于其天然但未变性状态时，其识别突变酵母蛋白二硫键异构酶的NCS/T和N-糖基化位点。这些结果表明，NXS/T基序的抗体识别是构象依赖性的，并表明NXS/T自发地采用抗体识别所必需的特定构象。这些特征可能与OT的已知结合特异性相关。
We aimed to identify antibodies that can recognize the Asn–Xaa–Ser/Thr(NXS/T) N-glycosylation site that guides oligosaccharyltransferase (OT) activity. We used synthetic Asn–Cys–Ser/Thr(NCS/T) tripeptides conjugated to bovine serum albumin to isolate single chain antibody fragments of a variable region (scFv) from the Griffin 1 phage antibody library. Although Ser and Thr have different side chains, the scFv proteins thus isolated bound to both NCS and NCT with Kdvalues of the order of 10−6M and accepted the substitution of the Cys residue with various amino acids, including Ala, Gly, and Val. However, these proteins recognized neither Asn–Pro–Ser/Thr nor non-NXS/T tripeptides. The scFv proteins recognized NCS/T and N-glycosylation site of mutant yeast protein disulfide isomerase when they were in their native but not denatured state. These results indicate that antibody recognition of the NXS/T motif is conformation dependent and suggest that NXS/T spontaneously adopts a specific conformation that is necessary for antibody recognition. These features are likely to correlate with the known binding specificity of OT.
影响因子：
3.9
作者：
Freeze, HH;Westphal, V
通讯作者：
Westphal, V
寡糖转移酶：内质网的一种复杂的多亚基酶。
DOI：
10.1006/bbrc.1999.1886
发表时间：
1999
期刊：
Biochemical and biophysical research communications.
影响因子：
--
作者：
Yan,Q;Lennarz,WJ
通讯作者：
Lennarz,WJ