Tokushima Institute, Otsuka Pharmaceutical Co., Ltd., Tokushima, Japan.
Proc Jpn Acad Ser B Phys Biol Sci. 2013;89(7):302-20. doi: 10.2183/pjab.89.302.
Through glycosphingolipid biochemical research, we developed two types of transcription technologies. One is a biochemical transfer of glycosphingolipids to peptides. The other is a physicochemical transfer of glycosphingolipids in silica gel to the surface of a plastic membrane. Using the first technology, we could prepare peptides which mimic the shapes of glycosphingolipid molecules by biopanning with a phage-displayed peptide library and anti-glycosphingolipid antibodies as templates. The peptides thus obtained showed biological properties and functions similar to those of the original glycosphingolipids, such as lectin binding, glycosidase modulation, inhibition of tumor metastasis and immune response against the original antigen glycosphingolipid, and we named them glyco-replica peptides. The results showed that the newly prepared peptides could be used effectively as a bio-recognition system and suggest that the glyco-replica peptides can be widely applied to therapeutic fields. Using the second technology, we could establish a functional lipidomics with a thin-layer chromatography-blot/matrix-assisted laser desorption ionization-time of flight mass spectrometry (TLC-Blot/MALDI-TOF MS) system. By transferring glycosphingolipids on a plastic membrane surface from a TLC plate, innovative biochemical approaches such as simple purification of individual glycosphingolipids, binding studies, and enzyme reactions could be developed. The combinations of these biochemical approaches and MALDI-TOF MS on the plastic membrane could provide new strategies for glycosphingolipid science and the field of lipidomics. In this review, typical applications of these two transfer technologies are introduced.(Communicated by Kunihiko SUZUKI, M.J.A.).
通过糖脂的生化研究,我们开发了两种转录技术。一种是糖脂的生化转移到肽。另一种是硅胶中糖脂的物理化学转移到塑料膜的表面。使用第一种技术,我们可以通过噬菌体展示肽库和抗糖脂抗体作为模板的生物淘选来制备模拟糖脂分子形状的肽。由此得到的肽表现出与原始糖脂相似的生物学性质和功能,如凝集素结合、糖苷酶调节、抑制肿瘤转移和对原始抗原糖脂的免疫反应,我们将它们命名为糖复制肽。结果表明,新制备的肽可有效用作生物识别系统,并表明糖复制肽可广泛应用于治疗领域。使用第二种技术,我们可以建立一个功能脂质组学与薄层色谱-斑点/基质辅助激光解吸电离飞行时间质谱(TLC-Blot/MALDI-TOF MS)系统。通过将糖脂从 TLC 板转移到塑料膜表面,可以开发出创新的生化方法,如单个糖脂的简单纯化、结合研究和酶反应。这些生化方法与塑料膜上的 MALDI-TOF MS 的组合可以为糖脂科学和脂质组学领域提供新的策略。在这篇综述中,介绍了这两种转移技术的典型应用。(由 Kunihiko SUZUKI,M.J.A. 传达)。