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首页> 外文期刊>Journal of Molecular Biology >Fucose depletion from human IgG1 oligosaccharide enhances binding enthalpy and association rate between IgG1 and FcgammaRIIIa.
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Fucose depletion from human IgG1 oligosaccharide enhances binding enthalpy and association rate between IgG1 and FcgammaRIIIa.

机译：从人IgG1寡糖中去除岩藻糖可增强IgG1与FcgRmIIIa之间的结合焓和缔合速率。

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Depletion of fucose from human IgG1 oligosaccharide improves its affinity for Fcgamma receptor IIIa (FcgammaRIIIa). This is the first case where a glycoform modification is shown to improve glycoprotein affinity for the receptors without carbohydrate-binding capacity, suggesting a novel glyco-engineering strategy to improve ligand-receptor binding. To address the mechanisms of affinity improvement by the fucose depletion, we used isothermal titration calorimetry (ITC) and biosensor analysis with surface plasmon resonance. ITC demonstrated that IgG1-FcgammaRIIIa binding was driven by favorable binding enthalpy (DeltaH) but opposed by unfavorable binding entropy change (DeltaS). Fucose depletion from IgG1 enhanced the favorable DeltaH, leading to the increase in the binding constant of IgG1 for the receptor by a factor of 20-30. The increase in the affinity was mainly attributed to an enhanced association rate. A triple amino acid substitution in IgG1, S298A/E333A/K334A, is also known to improve IgG1 affinity for FcgammaRIIIa. ITC demonstrated that the amino acid substitution attenuated the unfavorable DeltaS resulting in a three- to fourfold increase in the binding constant. The affinity enhancement by the amino acid substitution was due to a reduced dissociation rate. These results indicate that the mechanism of affinity improvement by the fucose depletion is quite distinct from that by the amino acid substitution. Defucosylated IgG1 exhibited higher antibody-dependent cellular cytotoxicity (ADCC) than S298A/E333A/K334A-IgG1, showing a correlation between IgG1 affinity for FcgammaRIIIa and ADCC. We also examined the effect of FcgammaRIIIa polymorphism (Val158/Phe158) on IgG1-FcgammaRIIIa binding. The Phe to Val substitution increased FcgammaRIIIa affinity for IgG1 in an enthalpy-driven manner with the reduced dissociation rate. These results together highlight the distinctive functional improvement of affinity by IgG1 defucosylation and suggest that engineering of non-interfacial monosaccharides can improve glycoprotein affinity for receptors via an enthalpy-driven and association rate-assisted mechanism.
机译：从人IgG1低聚糖中去除岩藻糖可改善其对Fcgamma受体IIIa（FcgammaRIIIa）的亲和力。这是第一种情况，显示糖型修饰可改善糖蛋白对受体的亲和力，而无糖结合能力，这表明可改善配体-受体结合的新型糖工程策略。为了解决由于岩藻糖耗竭而提高亲和力的机制，我们使用了等温滴定热量法（ITC）和具有表面等离子体共振的生物传感器分析。 ITC证明IgG1-FcgammaRIIIa的结合是由有利的结合焓（DeltaH）驱动的，但受到不利的结合熵变化（DeltaS）的反对。 IgG1的岩藻糖消耗增强了有利的DeltaH，导致IgG1与受体的结合常数增加了20-30倍。亲和力的增加主要归因于关联率的提高。 IgG1中的三氨基酸取代，即S298A / E333A / K334A，也可以改善IgG1对FcgRIIIa的亲和力。 ITC证明，氨基酸取代减弱了不利的DeltaS，导致结合常数增加了三到四倍。氨基酸取代增加的亲和力是由于解离速率降低。这些结果表明，通过岩藻糖耗竭改善亲和力的机理与通过氨基酸取代而改善亲和力的机理非常不同。与S298A / E333A / K334A-IgG1相比，去岩藻糖基化的IgG1表现出更高的抗体依赖性细胞毒性（ADCC），表明IgG1对FcgammaRIIIa和ADCC的亲和力之间存在相关性。我们还检查了FcgammaRIIIa多态性（Val158 / Phe158）对IgG1-FcgammaRIIIa结合的影响。从Phe到Val的置换以焓驱动方式增加了FcgammaRIIIa对IgG1的亲和力，且解离速率降低。这些结果共同突出了通过IgG1去岩藻糖基化显着改善亲和力的功能，并表明非界面单糖的工程化可以通过焓驱动和缔合速率辅助机制改善糖蛋白对受体的亲和力。

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