脂質と抗原提示タンパク質の相互作用の解明を目指した結合分子の合成と生物活性評価

我们合成了一系列新的溶血磷脂酰乙醇胺，它们是从小鼠胸腺中分离的NKT活化缩醛磷脂（pLPE）的衍生物。将人CD 1d-β 2 M固定在CM 5传感器芯片上，通过表面等离子体共振（SPR）技术检测了脂质与CD 1d的结合能力，并检测了合成的配体和其他固有配体与CD 1d的结合能力。首先使用标准配体KRN 7000建立动力学测量条件，然后确定合成和其他天然溶血磷脂的结合亲和力。所有溶血磷脂对hCD 1d固定化表面显示出中等的解离常数（KD = 85 - 253 nM）。使用蜡酸作为阴性对照，其显示与CD 1d修饰的表面没有结合亲和力。其中，在尾部具有双键或对氟苄基的磷脂比饱和或未取代的衍生物显示出略强的结合亲和力。除了乙烯基醚连接外，头尾基团的连接性在结合亲和力中没有显示出任何显著性。乙醇胺组的存在或不存在显示出结合亲和力的显著差异。另一方面，乙醇胺到胆碱的转化没有显示出任何结合活性差异。

We synthesized a novel series of lysophosphatidylethanolamine, which are derivatives of NKT activating plasmalogens (pLPE) isolated from mouse thymus. We have established SPR technique to examine the lipid-CD1d binding affinity and examined the binding affinity of the synthesized and some other intrinsic ligands with CD1d.Human CD1d-β2M was immobilized on CM5 sensor chip and ligands solution was passed through the modified surface for the binding analysis. The kinetic measurement conditions are established first using standard ligand KRN7000, then the binding affinities of the synthesized and other natural lysophospholipids. All the lysophospholipids showed moderate dissociation constants (KD = 85 -253 nM) to the hCD1d immobilized surface. Cerotic acid used as a negative control, which showed no binding affinity with CD1d modified surface. Among the phospholipids, which have either double bond or p-fluoro benzyl group in the tail, showed slightly stronger binding affinities than the saturated or unsubstituted derivatives. The connectivity of head-tail groups does not show any significance in the binding affinity except vinyl-ether connection. Absence or presence of ethanolamine group showed significant difference in the binding affinity. On the other hand, ethanolamine to choline transformation did not show any binding activity difference.