广谱中和抗体 PGT121 通过识别 HIV-1 gp120 V3 基部和多个周围聚糖,变构调节 CD4 结合。

Broadly neutralizing antibody PGT121 allosterically modulates CD4 binding via recognition of the HIV-1 gp120 V3 base and multiple surrounding glycans.

机构信息

Department of Integrative Structural and Computational Biology, The Scripps Research Institute, La Jolla, California, USA.

出版信息

PLoS Pathog. 2013;9(5):e1003342. doi: 10.1371/journal.ppat.1003342. Epub 2013 May 2.

Abstract

New broad and potent neutralizing HIV-1 antibodies have recently been described that are largely dependent on the gp120 N332 glycan for Env recognition. Members of the PGT121 family of antibodies, isolated from an African donor, neutralize ∼70% of circulating isolates with a median IC50 less than 0.05 µg ml(-1). Here, we show that three family members, PGT121, PGT122 and PGT123, have very similar crystal structures. A long 24-residue HCDR3 divides the antibody binding site into two functional surfaces, consisting of an open face, formed by the heavy chain CDRs, and an elongated face, formed by LCDR1, LCDR3 and the tip of the HCDR3. Alanine scanning mutagenesis of the antibody paratope reveals a crucial role in neutralization for residues on the elongated face, whereas the open face, which accommodates a complex biantennary glycan in the PGT121 structure, appears to play a more secondary role. Negative-stain EM reconstructions of an engineered recombinant Env gp140 trimer (SOSIP.664) reveal that PGT122 interacts with the gp120 outer domain at a more vertical angle with respect to the top surface of the spike than the previously characterized antibody PGT128, which is also dependent on the N332 glycan. We then used ITC and FACS to demonstrate that the PGT121 antibodies inhibit CD4 binding to gp120 despite the epitope being distal from the CD4 binding site. Together, these structural, functional and biophysical results suggest that the PGT121 antibodies may interfere with Env receptor engagement by an allosteric mechanism in which key structural elements, such as the V3 base, the N332 oligomannose glycan and surrounding glycans, including a putative V1/V2 complex biantennary glycan, are conformationally constrained.

摘要

最近描述了新的广谱和强效中和 HIV-1 抗体,这些抗体在很大程度上依赖于 gp120 N332 聚糖来识别 Env。从一名非洲供体中分离出的 PGT121 抗体家族成员,可中和约 70%的循环分离株,其半数抑制浓度(IC50)小于 0.05 µg ml(-1)。在这里,我们展示了三个家族成员 PGT121、PGT122 和 PGT123 具有非常相似的晶体结构。一个长的 24 个残基的 HCDR3 将抗体结合位点分为两个功能表面,由重链 CDR 组成的开放面和由 LCDR1、LCDR3 和 HCDR3 尖端组成的伸长面。对抗体变构区的丙氨酸扫描诱变揭示了在中和作用中,伸长面的残基起着至关重要的作用,而在 PGT121 结构中容纳复杂的双天线聚糖的开放面则起着次要作用。对工程重组 Env gp140 三聚体(SOSIP.664)的负染 EM 重建显示,PGT122 与 gp120 外域的相互作用相对于刺突的顶面呈更垂直的角度,而之前表征的依赖于 N332 聚糖的抗体 PGT128 则呈更水平的角度。然后,我们使用 ITC 和 FACS 证明,尽管表位远离 CD4 结合位点,但 PGT121 抗体可抑制 CD4 与 gp120 的结合。综上所述,这些结构、功能和生物物理结果表明,PGT121 抗体可能通过变构机制干扰 Env 受体的结合,在这种机制中,关键结构元件,如 V3 基部、N332 寡甘露糖聚糖和周围聚糖,包括一个假定的 V1/V2 复杂双天线聚糖,被构象约束。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0535/3642082/fa2658fc6a65/ppat.1003342.g001.jpg

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