How altering the modular architecture affects aspects of lectin activity: case study on human galectin-1.
How altering the modular architecture affects aspects of lectin activity: case study on human galectin-1.
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DOI：
10.1093/glycob/cwz034
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发表时间：
2019-05
期刊：
影响因子：
4.3
通讯作者：
Tanja J. Kutzner;A. Gabba;Forrest G. FitzGerald;N. Shilova;Gabriel García Caballero;Anna‐Kristin Ludwig;J. C. Manning;C. Knospe;H. Kaltner;F. Sinowatz;P. Murphy;M. Cudic;N. Bovin;H. Gabius
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文献类型：
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作者：
Tanja J. Kutzner;A. Gabba;Forrest G. FitzGerald;N. Shilova;Gabriel García Caballero;Anna‐Kristin Ludwig;J. C. Manning;C. Knospe;H. Kaltner;F. Sinowatz;P. Murphy;M. Cudic;N. Bovin;H. Gabius
在许多（病理）生理过程中涉及聚糖-蛋白质识别的发现正将注意力引导到探索糖受体的基本结构方面的重要性而不仅仅是聚糖特异性，即，出现不同类型的模块化架构。为了追踪用于定义人凝集素中的设计-功能关系的线索，凝集素的结构单元已被用作工程化定制野生型蛋白变体的源材料。它们的可用性有助于对所述目标进行比较分析。以粘附/生长调节性人半乳糖凝集素-1为例，通过使用聚糖复杂性程度增加的三种测定系统来说明评价其设计变化（此处，从非共价缔合结构域的同源二聚体到（i）接头连接的二聚体和四聚体以及（ii）半乳糖凝集素-3样蛋白）如何影响活性的策略。尽管用两种同源二糖的量热法和用647（糖）化合物的阵列测试没有揭示重大变化，但呈现天然糖组复杂性的组织切片的半乳糖凝集素组织化学染色谱揭示了野生型和接头连接的同源寡聚物之间以及半乳糖凝集素-3样变体和野生型半乳糖凝集素-3之间的细胞类型阳性差异，同一位点的强度水平和对二价糖化合物抑制的敏感性。这些结果强调了记录方法的优势。此外，他们给出了方向，以继续（i）将其应用扩展到该凝集素家族的其他成员，特别是半乳糖凝集素-3，以及（ii）然后分析结构改变对细胞表面晶格形成的影响，并系统地进行生物信号传导，考虑变体的转化医学潜力。
Discoveries on involvement of glycan-protein recognition in many (patho)physiological processes are directing attention to exploring the significance of a fundamental structural aspect of sugar receptors beyond glycan specificity, i.e., occurrence of distinct types of modular architecture. In order to trace clues for defining design-functionality relationships in human lectins, a lectin's structural unit has been used as source material for engineering custom-made variants of the wild-type protein. Their availability facilitates comparative analysis toward the stated aim. With adhesion/growth-regulatory human galectin-1 as example, the strategy of evaluating how changes of its design (here, from the homodimer of non-covalently associated domains to (i) linker-connected di- and tetramers and (ii) a galectin-3-like protein) affect activity is illustrated by using three assay systems of increasing degree of glycan complexity. Whereas calorimetry with two cognate disaccharides and array testing with 647 (glyco)compounds disclosed no major changes, galectin histochemical staining profiles of tissue sections that present natural glycome complexity revealed differences between wild-type and linker-connected homo-oligomers as well as between the galectin-3-like variant and wild-type galectin-3 for cell-type positivity, level of intensity at the same site and susceptibility for inhibition by a bivalent glycocompound. These results underscore the strength of the documented approach. Moreover, they give direction to proceed to (i) extending its application to other members of this lectin family, especially galectin-3 and (ii) then analyzing impact of architectural alterations on cell surface lattice formation and ensuing biosignaling systematically, considering the variants' potential for translational medicine.