BAFF/BLyS受体3包含一个最小的肿瘤坏死因子受体样模块,该模块编码一个高度专一的配体结合位点。
BAFF/BLyS receptor 3 comprises a minimal TNF receptor-like module that encodes a highly focused ligand-binding site.
作者信息
Gordon Nathaniel C, Pan Borlan, Hymowitz Sarah G, Yin JianPing, Kelley Robert F, Cochran Andrea G, Yan Minhong, Dixit Vishva M, Fairbrother Wayne J, Starovasnik Melissa A
机构信息
Department of Protein Engineering, Genentech, Inc., One DNA Way, South San Francisco, California 94080, USA.
出版信息
Biochemistry. 2003 May 27;42(20):5977-83. doi: 10.1021/bi034017g.
BAFF/BLyS, a member of the tumor necrosis family (TNF) superfamily of ligands, is a crucial survival factor for B cells. BAFF binds three receptors, TACI, BCMA, and BR3, with signaling through BR3 being essential for promoting B cell function. Typical TNF receptor (TNFR) family members bind their cognate ligands through interactions with two cysteine-rich domains (CRDs). However, the extracellular domain (ECD) of BR3 consists of only a partial CRD, with cysteine spacing distinct from other modules described previously. Herein, we report the solution structure of the BR3 ECD. A core region of only 19 residues adopts a stable structure in solution. The BR3 fold is analogous to the first half of a canonical TNFR CRD but is stabilized by an additional noncanonical disulfide bond. BAFF-binding determinants were identified by shotgun alanine-scanning mutagenesis of the BR3 ECD expressed on phage. Several of the key BAFF-binding residues are presented from a beta-turn that we have shown previously to be sufficient for ligand binding when transferred to a structured beta-hairpin scaffold [Kayagaki, N., Yan, M., Seshasayee, D., Wang, H., Lee, W., French, D. M., Grewal, I. S., Cochran, A. G., Gordon, N. C., Yin, J., Starovasnik, M. A, and Dixit, V. M. (2002) Immunity 10, 515-524]. Outside of the turn, mutagenesis identifies additional hydrophobic contacts that enhance the BAFF-BR3 interaction. The crystal structure of the minimal hairpin peptide, bhpBR3, in complex with BAFF reveals intimate packing of the six-residue BR3 turn into a cavity on the ligand surface. Thus, BR3 binds BAFF through a highly focused interaction site, unprecedented in the TNFR family.
BAFF/BLyS是肿瘤坏死因子(TNF)超家族配体成员,是B细胞的关键存活因子。BAFF与三种受体TACI、BCMA和BR3结合,通过BR3发出的信号对于促进B细胞功能至关重要。典型的TNF受体(TNFR)家族成员通过与两个富含半胱氨酸的结构域(CRD)相互作用来结合其同源配体。然而,BR3的细胞外结构域(ECD)仅由部分CRD组成,其半胱氨酸间距与先前描述的其他模块不同。在此,我们报道了BR3 ECD的溶液结构。仅19个残基的核心区域在溶液中采用稳定结构。BR3折叠类似于典型TNFR CRD的前半部分,但通过额外的非典型二硫键得以稳定。通过对噬菌体上表达的BR3 ECD进行鸟枪法丙氨酸扫描诱变来鉴定BAFF结合决定簇。几个关键的BAFF结合残基来自一个β转角,我们之前已经表明,当转移到结构化的β发夹支架上时,该转角足以实现配体结合[Kayagaki, N., Yan, M., Seshasayee, D., Wang, H., Lee, W., French, D. M., Grewal, I. S., Cochran, A. G., Gordon, N. C., Yin, J., Starovasnik, M. A, and Dixit, V. M. (2002) Immunity 10, 515 - 524]。在转角之外,诱变鉴定出了增强BAFF - BR3相互作用的其他疏水接触。与BAFF复合的最小发夹肽bhpBR3的晶体结构揭示了六残基BR3转角紧密堆积在配体表面的一个腔中。因此,BR3通过TNFR家族中前所未有的高度集中的相互作用位点结合BAFF。