McCarter Calvin, Kletter Doron, Tang Huiyuan, Partyka Katie, Ma Yinjiao, Singh Sudhir, Yadav Jessica, Bern Marshall, Haab Brian B
Van Andel Research Institute, Grand Rapids, MI 49503, USA.
Palo Alto Research Center, Palo Alto, CA, USA.
Proteomics Clin Appl. 2013 Oct;7(9-10):632-41. doi: 10.1002/prca.201300069. Epub 2013 Sep 13.
Lectins are valuable tools for detecting specific glycans in biological samples, but the interpretation of the measurements can be ambiguous due to the complexities of lectin specificities. Here, we present an approach to improve the accuracy of interpretation by converting lectin measurements into quantitative predictions of the presence of various glycan motifs.
The conversion relies on a database of analyzed glycan array data that provides information on the specificities of the lectins for each of the motifs. We tested the method using measurements of lectin binding to glycans on glycan arrays and then applied the method to predicting motifs on the protein mucin 1 (MUC1) expressed in eight different pancreatic cancer cell lines.
The combined measurements from several lectins were more accurate than individual measurements for predicting the presence or absence of motifs on arrayed glycans. The analysis of MUC1 revealed that each cell line expressed a unique pattern of glycoforms, and that the glycoforms significantly differed between MUC1 collected from conditioned media and MUC1 collected from cell lysates.
This new method could provide more accurate analyses of glycans in biological sample and make the use of lectins more practical and effective for a broad range of researchers.
凝集素是检测生物样品中特定聚糖的重要工具,但由于凝集素特异性的复杂性,测量结果的解读可能存在歧义。在此,我们提出一种方法,通过将凝集素测量结果转化为各种聚糖基序存在的定量预测,来提高解读的准确性。
这种转化依赖于一个分析过的聚糖阵列数据数据库,该数据库提供了凝集素对每个基序特异性的信息。我们使用凝集素与聚糖阵列上聚糖结合的测量结果来测试该方法,然后将该方法应用于预测八种不同胰腺癌细胞系中表达的蛋白质粘蛋白1(MUC1)上的基序。
对于预测阵列上聚糖基序的存在与否,几种凝集素的联合测量比单独测量更准确。对MUC1的分析表明,每个细胞系表达独特的糖型模式,并且从条件培养基中收集的MUC1与从细胞裂解物中收集的MUC1之间的糖型存在显著差异。
这种新方法可以对生物样品中的聚糖进行更准确的分析,并使凝集素的使用对广大研究人员来说更实用、更有效。