NMR INVESTIGATION OF HUMAN CELL SURFACE RECEPTOR CD58: IMMUNOLOGY

人类细胞表面受体 CD58 的 NMR 研究：免疫学

• 批准号: 6355184
• 负责人: ZHEN-YU JIM SUN
• 金额: $ 0.11万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-05-01 至 2001-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6355184
• 关键词: biological products; biomedical resource; proteins; technology /technique

The goals of this research project are to determine the NMR solution structure of the adhesion domain of human cell surface receptor CD58, to study its interaction with the counter receptor CD2 and to develop a general strategy for producing a glycan-free form of glycoproteins without loss of function. The major result of this research project during 1998 is the successful NMR structural determination of a soluble mutant form of the CD58 adhesion domain, obtained through a structure-based rationalized design employing apolar-to-polar mutations of surface residues in functionally-irrelevant epitopes. The success of this structure-based approach was demonstrated through the E-coli expression of this functional 11 kDa adhesion domain extracted from the heavily glycosylated 55 kDa human CD58 ectodomain. The solution structure of the CD58 adhesion domain was subsequently determined using the NOESY. Other critical data were obtained from the Varian 750 MHz spectrometer at CMR, where we also studied via NMR the binding to its counter-receptor CD2. It has been shown that the CD58 adhesion domain adopts the immunoglobulin variable domain fold and consists of nine beta strands forming two parallel beta sheets. The new structural information supports a "hand-shake" model of CD2/CD58 interaction involving the GFCCU faces of both CD2 and CD58 adhesion domains. The region around the CC'Ioop of CD58 is most likely responsible for binding specificity while the negatively charged residues around the concave region near FG loop appear to contribute mainly to guiding the orientation during the docking of the two receptors. The structural information obtained thus far leads to promising experiments aimed at studying the solution structure of the complex formed between the two adhesion domains of CD58 and CD2. Due to the increased complexity of the CD2/CD58 binding complex structure, the experimental data from the 750 Nfflz spectrometer will have an even greater significance.

这项研究项目的目标是确定核磁共振 人细胞表面粘附域的溶液结构 受体CD58，研究其与拮抗受体CD2的相互作用 并开发一种通用策略来生产一种无糖形式的 不丧失功能的糖蛋白。这一点的主要结果是 1998年的研究项目是成功的核磁共振结构 CD58粘附域的可溶性突变形式的测定， 通过采用基于结构的合理化设计获得 细菌表面残基的非极-极突变 功能无关的表位。这种基于结构的成功 通过对该基因的原核表达，验证了该方法 从重组腺病毒中提取功能性11 kDa粘附区 糖基化55 kDa人CD58胞外结构域。的解结构 随后使用NOESY测定CD58粘附域。 其他关键数据是从瓦里安750兆赫光谱仪获得的。 在CMR，我们也通过核磁共振研究了与其结合 受体拮抗剂CD2。已有研究表明，CD58的粘附域 采用免疫球蛋白可变区折叠，由九个 形成两个平行的β片层的β链。新的结构 信息支持CD2/CD58交互的“握手”模式 涉及CD2和CD58粘附域的GFCCU表面。这个 CD58基因CC‘oop周围的区域最有可能是 结合的特异性而周围的带负电荷的残基 FG循环附近的凹面区域似乎主要有助于引导 在两个受体对接过程中的定向。结构性的 到目前为止获得的信息导致了旨在 研究两者之间形成的络合物的溶液结构 CD58和CD2的粘附域。由于……的复杂性增加 CD2/CD58结合络合物结构的实验数据 750nfflz光谱仪将具有更大的意义。