MHCL RESTRICTED TCR RECOGNITION--STRUCTURAL BASIS

MHCL 限制性 TCR 识别——结构基础

• 批准号: 2472505
• 负责人: JIA-HUAI WANG
• 金额: $ 22.31万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-01-01 至 2002-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2472505
• 关键词: T cell receptor; Vesiculovirus; X ray crystallography; antireceptor antibody; histocompatibility antigens; immunoglobulin structure; receptor binding; virus antigen

DESCRIPTION (Adapted from Investigator's abstract): Soluble T cell receptors (TCRs) have been engineered for secretion by Lec3.2.8.1 CHO cells which synthesize homogeneous gylcans; a leucine zipper is used to facilitate a-b chain association. Two mouse TCRs, N15 and N26, specific for vesicular stomatitis virus octapeptide (VSV8) in the context of the Kb class I molecule, have been expressed by this method. Crystals of N15 and N26 complexed with the Fab of an anti-Cb monoclonal antibody (FabH57) and of the N15-VSV8/Kb and N26-VSV8/Kb complexes have been obtained. The structure of the N15-FabN15 complex has been solved to 2.8 A resolution by molecular replacement in conjunction with selenomethionine/MAD phasing. The structure of the N15 TCR will be compared with that of other Ig superfamily members, including antibodies, CD4, VCAM-1 and CD2. By comparing each of two complexes in the asymmetric unit of N15-FabH57 crystals, it may be possible to ascertain whether there is intrinsic mobility between V-V, V-C and/or C-C domains of the TCR. The structure of the N15-VSV8/Kb complex will be solved by molecular replacement using crystals which diffract to 3 A resolution. The role of individual CDRs in peptide/MHC recognition will be determined and any induced fit upon antigen binding analyzed by comparing the structure of the CDRs in the TCR-VSV8/Kb versus TCR-Fab complex. Structure analysis of the N26 TCR complexed with VSV8/Kb will also be carried out in order to determine how two different TCRs bind the same peptide/MHC ligand. This should establish whether the docking mode is fixed or variable and whether there are restricted interactions between specific CDRs and MHC versus peptide residues.

描述(改编自研究人员摘要)：可溶性T细胞 受体(TCR)已被设计用于Lec3.2.8.1 CHO细胞的分泌 它合成了均匀的糖罐；使用亮氨酸拉链来促进 A-b链缔合。两个小鼠TCR，n15和n26，专用于水泡 KB类中的口炎病毒八肽(VSV8) 分子，已经用这种方法表达了。N15和N26的晶体 与抗CB单抗Fab(FabH57)和抗CB单抗Fab络合 得到了N15-VSV8/Kb和N26-VSV8/Kb络合物。的结构 N15-FabN15络合物已被分子分解到2.8A分辨率 结合硒蛋氨酸/MAD分期进行替换。该结构 将N15 TCR与其他免疫球蛋白超家族成员的基因进行比较， 包括抗体、CD4、VCAM-1和CD2。通过比较两个 在不对称单元的N15-FabH57晶体中形成络合物，这是可能的 确定V-V、V-C和/或C-C之间是否存在本征迁移率 TCR的域。N15-VSV8/Kb复合体的结构将被解决 通过使用衍射到3A分辨率的晶体进行分子置换。 将确定单个CDR在多肽/MHC识别中的作用 以及通过比较结构来分析任何与抗原结合相匹配的诱导 TCR-VSV8/Kb与TCR-Fab复合体中CDRs的比例。结构分析 还将进行与VSV8/Kb复合的n26 TCR，以便 确定两个不同的TCR如何结合相同的多肽/MHC配体。这 应该确定对接模式是固定的还是可变的，以及 特定的CDR和MHC之间的相互作用受到限制 多肽残留物。