Chemical and structural analysis of an antibody folding intermediate trapped during glycan biosynthesis.
Chemical and structural analysis of an antibody folding intermediate trapped during glycan biosynthesis.
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DOI：
10.1021/ja306068g
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发表时间：
2012-10-24
影响因子：
15
通讯作者：
Crispin, Max
中科院分区：
文献类型：
--
作者：
Bowden, Thomas A.;Baruah, Kavitha;Coles, Charlotte H.;Harvey, David J.;Yu, Xiaojie;Song, Byeong-Doo;Stuart, David I.;Aricescu, A. Radu;Scanlan, Christopher N.;Jones, E. Yvonne;Crispin, Max
人IgG Fc糖基化调节免疫效应子功能，如抗体依赖性细胞毒性和吞噬作用。因此，Fc聚糖的工程改造使得能够微调单克隆抗体的治疗性质。Fc的N-连接聚糖通常为复合型，沿Cγ2结构域的蛋白质表面沿着形成非共价相互作用网络。在这里，我们操纵的哺乳动物聚糖加工途径，以陷阱IgG 1 Fc在成熟的顺序阶段，从寡甘露糖-杂交-复合型聚糖，并表明，Fc结构稳定的聚糖从其杂交-复合型状态的过渡。这种混合型中间体的X射线晶体学分析表明，N-连接的聚糖在成熟后发生构象变化，包括三甘露糖核心内的翻转。该中间体的晶体结构揭示了抗体生物发生的分子基础，并为治疗性抗体的蛋白质-聚糖界面的结构指导工程提供了模板。
Human IgG Fc glycosylation modulates immunological effector functions such as antibody-dependent cellular cytotoxicity and phagocytosis. Engineering of Fc glycans therefore enables fine-tuning of the therapeutic properties of monoclonal antibodies. The N-linked glycans of Fc are typically complex-type, forming a network of noncovalent interactions along the protein surface of the Cγ2 domain. Here, we manipulate the mammalian glycan-processing pathway to trap IgG1 Fc at sequential stages of maturation, from oligomannose- to hybrid- to complex-type glycans, and show that the Fc is structurally stabilized following the transition of glycans from their hybrid- to complex-type state. X-ray crystallographic analysis of this hybrid-type intermediate reveals that N-linked glycans undergo conformational changes upon maturation, including a flip within the trimannosyl core. Our crystal structure of this intermediate reveals a molecular basis for antibody biogenesis and provides a template for the structure-guided engineering of the protein–glycan interface of therapeutic antibodies.
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影响因子：
4
作者：
Borrok, M. Jack;Jung, Sang Taek;Kang, Tae Hyun;Monzingo, Arthur F.;Georgiou, George
通讯作者：
Georgiou, George
影响因子：
4.3
作者：
Butler, M;Quelhas, D;Rudd, PM
通讯作者：
Rudd, PM
DOI：
10.1107/s0907444904019158
发表时间：
2004-12-01
影响因子：
2.2
作者：
Emsley, P;Cowtan, K
通讯作者：
Cowtan, K
影响因子：
3
作者：
Dalziel, M;McFarlane, I;Axford, JS
通讯作者：
Axford, JS
人IgG1分子上存在的寡聚物种的血清清除率增加。
影响因子：
5.3
作者：
Alessandri L;Ouellette D;Acquah A;Rieser M;Leblond D;Saltarelli M;Radziejewski C;Fujimori T;Correia I
通讯作者：
Correia I