Kopitz Jürgen, Vértesy Sabine, André Sabine, Fiedler Sabine, Schnölzer Martina, Gabius Hans-Joachim
Abteilung für Angewandte Tumorbiologie, Zentrum Pathologie, Klinikum der Ruprecht-Karls-Universität, Im Neuenheimer Feld 224, 69120 Heidelberg, Germany.
Institut für Physiologische Chemie, Tierärztliche Fakultät, Ludwig-Maximilians-Universität, Veterinärstraße 13, 80539 München, Germany.
Biochimie. 2014 Sep;104:90-9. doi: 10.1016/j.biochi.2014.05.010. Epub 2014 Jun 6.
Many human proteins have a modular design with receptor and structural domains. Using adhesion/growth-regulatory galectin-3 as model, we describe an interdisciplinary strategy to define the functional significance of its tail established by nine non-triple helical collagen-like repeats (I-IX) and the N-terminal peptide. Genetic engineering with sophisticated mass spectrometric product analysis provided the tools for biotesting, i.e. eight protein variants with different degrees of tail truncation. Evidently,various aspects of galectin-3 activity (cis binding and cell bridging) are affected by tail shortening in a different manner. Thus, this combined approach reveals an unsuspected complexity of structure-function relationship, encouraging further application beyond this chimera-type galectin.
许多人类蛋白质具有模块化设计，包含受体和结构域。以黏附/生长调节半乳糖凝集素-3为模型，我们描述了一种跨学科策略，以确定由九个非三螺旋胶原样重复序列（I-IX）和N端肽段构成的其尾部的功能意义。结合精密质谱产物分析的基因工程为生物测试提供了工具，即产生了八个具有不同程度尾部截短的蛋白质变体。显然，半乳糖凝集素-3活性的各个方面（顺式结合和细胞桥接）受尾部缩短的影响方式不同。因此，这种联合方法揭示了结构-功能关系中未曾预料到的复杂性，促使该方法在这种嵌合型半乳糖凝集素之外得到进一步应用。
