Site-Selective Chemoenzymatic Modification on the Core Fucose of an Antibody Enhances Its Fcγ Receptor Affinity and ADCC Activity.
Site-Selective Chemoenzymatic Modification on the Core Fucose of an Antibody Enhances Its Fcγ Receptor Affinity and ADCC Activity.
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对抗体核心岩藻糖进行位点选择性化学酶修饰可增强其 Fcγ 受体亲和力和 ADCC 活性。
DOI：
10.1021/jacs.1c03174
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发表时间：
2021-05-26
影响因子：
15
通讯作者：
Wang LX
中科院分区：
文献类型：
--
作者：
Li C;Chong G;Zong G;Knorr DA;Bournazos S;Aytenfisu AH;Henry GK;Ravetch JV;MacKerell AD Jr;Wang LX
Fc糖基化深刻地影响抗体的效应子功能，并且通常决定抗体的促炎或抗炎活性。已充分确定抗体Fc结构域N-聚糖的核心岩藻糖基化显著降低其对FcγRIIIa受体的亲和力和抗体依赖性细胞毒性（ADCC）。先前的结构研究表明，核心岩藻糖的存在显著降低了Fc和受体N-聚糖之间独特且有利的碳水化合物-碳水化合物相互作用，导致亲和力降低。我们在此报告，与天然核心岩藻糖相比，核心岩藻糖上的特殊位点特异性修饰可显著增强抗体对FcγRIIIa的亲和力。通过用新型岩藻糖苷酶突变体进行酶促岩藻糖基转移实现了位点选择性修饰，该突变体显示能够使用修饰的α-岩藻糖基氟化物作为供体底物。我们发现，用6-叠氮或6-羟基-1-岩藻糖（1-半乳糖）替换核心1-岩藻糖显著增强了抗体对FcγRIIIa受体的亲和力，并显著增加了ADCC活性。为了理解修饰的岩藻糖介导的亲和力增强的机制，我们进行了分子动力学模拟。我们的数据显示，选择性核心-岩藻糖修饰增加了Fc和Fc受体之间的聚糖接触数量，显示了独特的碳水化合物-碳水化合物相互作用在实现抗体的高FcγRIIIa亲和力和ADCC活性方面的重要性。因此，直接位点选择性修饰将核心岩藻糖的不利影响转化为促进碳水化合物-碳水化合物相互作用的有利力。
Fc glycosylation profoundly impacts the effector functions of antibodies and often dictates an antibody’s pro- or anti-inflammatory activities. It is well established that core fucosylation of the Fc domain N-glycans of an antibody significantly reduces its affinity for FcγRIIIa receptors and antibody-dependent cellular cytotoxicity (ADCC). Previous structural studies have suggested that the presence of a core fucose remarkably decreases the unique and favorable carbohydrate–carbohydrate interactions between the Fc and the receptor N-glycans, leading to reduced affinity. We report here that in contrast to natural core fucose, special site-specific modification on the core fucose could dramatically enhance the affinity of an antibody for FcγRIIIa. The site-selective modification was achieved through an enzymatic transfucosylation with a novel fucosidase mutant, which was shown to be able to use modified α-fucosyl fluoride as the donor substrate. We found that replacement of the core l-fucose with 6-azide- or 6-hydroxy-l-fucose (l-galactose) significantly enhanced the antibody’s affinity for FcγRIIIa receptors and substantially increased the ADCC activity. To understand the mechanism of the modified fucose-mediated affinity enhancement, we performed molecular dynamics simulations. Our data revealed that the number of glycan contacts between the Fc and the Fc receptor was increased by the selective core-fucose modifications, showing the importance of unique carbohydrate–carbohydrate interactions in achieving high FcγRIIIa affinity and ADCC activity of antibodies. Thus, the direct site-selective modification turns the adverse effect of the core fucose into a favorable force to promote the carbohydrate–carbohydrate interactions.
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影响因子：
7.4
作者：
Boero S;Morabito A;Banelli B;Cardinali B;Dozin B;Lunardi G;Piccioli P;Lastraioli S;Carosio R;Salvi S;Levaggi A;Poggio F;D'Alonzo A;Romani M;Del Mastro L;Poggi A;Pistillo MP
通讯作者：
Pistillo MP
影响因子：
4
作者：
Falconer DJ;Subedi GP;Marcella AM;Barb AW
通讯作者：
Barb AW
影响因子：
46.9
作者：
Li, HJ;Sethuraman, N;Gerngross, TU
通讯作者：
Gerngross, TU
影响因子：
4.7
作者：
Cumnock, Katherine;Tully, Timothy;Jacobson, Fredric
通讯作者：
Jacobson, Fredric
影响因子：
5.5
作者：
Best, Robert B.;Zhu, Xiao;Shim, Jihyun;Lopes, Pedro E. M.;Mittal, Jeetain;Feig, Michael;MacKerell, Alexander D., Jr.
通讯作者：
MacKerell, Alexander D., Jr.