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Structural characterization of a low molecular weight receptor for peanut agglutinin in murine lymphocytes

机译:鼠淋巴细胞中花生凝集素低分子量受体的结构表征

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A novel low molecular weight (LMW) receptor for the lectin peanut agglutinin (PNA) was characterized and purified from murine thymocytes and peripheral lymphocytes in the spleen. Preliminary binding assays with the fluoresceinated lectin demonstrated that the majority of cortical thymocytes, and only a fraction of lymphocytes in the splenic white pulp, as well as other peripheral lymphoid organs were PNA-positive. This positivity was selectively inhibited by the preferred PNA disaccharide ligand (Galβ1,3GalNAc), indicating the specificity of the binding. PNA receptors were purified from thymocytes and splenocytes by affinity chroma-tography on a PNA-agarose column, and their structural characteristics assessed by treatments with endoglycosidases and alkaline borohydride and analysis by two-dimensional (2-D) gels. Comparisons based on 2-D gels of glycosylated and deglycosylated forms were consistent with the thymic and splenic receptors, sharing a common 21 kDa polypeptide backbone, which is subjected to differential post-translational N-linked glycosylations with one (thymic PNA receptor) or two and/or three (splenic PNA receptor) complex-type glycan units of distinct structures. Analyses of amino acid and carbohydrate compositions of the intact receptors confirmed these observations and revealed a comparatively high level of sialic acid residues in the splenic PNA receptor.
机译:凝集素花生凝集素(PNA)的新型低分子量(LMW)受体的特点是从脾脏中的鼠胸腺细胞和外周淋巴细胞中纯化得到。荧光素凝集素的初步结合试验表明,脾脏白髓中的大部分皮质胸腺细胞,以及仅一部分淋巴细胞以及其他外周淋巴器官均为PNA阳性。该阳性反应被优选的PNA二糖配体(Galβ1,3GalNAc)选择性抑制,表明结合的特异性。通过在PNA-琼脂糖柱上进行亲和层析从胸腺细胞和脾细胞中纯化PNA受体,并通过用糖苷内切酶和碱性硼氢化物处理并通过二维(2-D)凝胶分析来评估其结构特征。基于糖基化和去糖基化形式的2-D凝胶进行的比较与胸腺和脾脏受体一致,共享一个共同的21 kDa多肽骨架,该骨架与一个(胸腺PNA受体)或两个经历不同的翻译后N-联糖基化和/或三个(脾PNA受体)结构不同的复杂型聚糖单元。对完整受体的氨基酸和碳水化合物组成的分析证实了这些观察结果,并揭示了脾脏PNA受体中唾液酸残基的水平相对较高。

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