Dynamic interplay between catalytic and lectin domains of GalNAc-transferases modulates protein O-glycosylation.
Dynamic interplay between catalytic and lectin domains of GalNAc-transferases modulates protein O-glycosylation.
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DOI：
10.1038/ncomms7937
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发表时间：
2015-05-05
影响因子：
16.6
通讯作者：
Hurtado-Guerrero, Ramon
中科院分区：
文献类型：
--
作者：
Lira-Navarrete, Erandi;de las Rivas, Matilde;Companon, Ismael;Carmen Pallares, Maria;Kong, Yun;Iglesias-Fernandez, Javier;Bernardes, Goncalo J. L.;Peregrina, Jesus M.;Rovira, Carme;Bernado, Pau;Bruscolini, Pierpaolo;Clausen, Henrik;Lostao, Anabel;Corzana, Francisco;Hurtado-Guerrero, Ramon
蛋白o -糖基化由多肽galnac -转移酶（GalNAc-Ts）控制，GalNAc-Ts具有独特的催化和凝集素结构域。galac - ts的凝集素结构域如何促进糖肽特异性和催化的潜在分子基础尚不清楚。在这里，我们提出了GalNAc-T2与糖肽配合物的第一个晶体结构，与增强的采样分子动力学模拟一起证明了凝集素结构域使糖肽的自由受体位点结合到催化结构域的合作机制。原子力显微镜和小角度x射线散射实验进一步揭示了GalNAc-T2的动态构象景观，以及高效催化所需的紧凑结构的突出作用。我们的模型表明GalNAc-T2的活性谱由构象异质性决定，并依赖于位于催化和凝集素结构域之间的柔性连接体。我们的研究结果也揭示了GalNAc-Ts如何用o -聚糖产生密集的蛋白质修饰。多肽galnac转移酶用密集的o -聚糖阵列修饰蛋白质，在粘蛋白的情况下，o -聚糖对于它们的屏障功能是必不可少的。在这里，作者提出了全面的结构研究，揭示了允许GalNAc-T2有效地进行密集o糖基化的分子属性。
Protein O-glycosylation is controlled by polypeptide GalNAc-transferases (GalNAc-Ts) that uniquely feature both a catalytic and lectin domain. The underlying molecular basis of how the lectin domains of GalNAc-Ts contribute to glycopeptide specificity and catalysis remains unclear. Here we present the first crystal structures of complexes of GalNAc-T2 with glycopeptides that together with enhanced sampling molecular dynamics simulations demonstrate a cooperative mechanism by which the lectin domain enables free acceptor sites binding of glycopeptides into the catalytic domain. Atomic force microscopy and small-angle X-ray scattering experiments further reveal a dynamic conformational landscape of GalNAc-T2 and a prominent role of compact structures that are both required for efficient catalysis. Our model indicates that the activity profile of GalNAc-T2 is dictated by conformational heterogeneity and relies on a flexible linker located between the catalytic and the lectin domains. Our results also shed light on how GalNAc-Ts generate dense decoration of proteins with O-glycans. Polypeptide GalNAc-transferases decorate proteins with dense arrays of O-glycans, which in the case of mucins are essential for their barrier functions. Here the authors present comprehensive structural studies that shed light on the molecular attributes that allow GalNAc-T2 to efficiently carry out dense O-glycosylation.
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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
DOI：
10.1073/pnas.202427399
发表时间：
2002-10-01
影响因子：
11.1
作者：
Laio, A;Parrinello, M
通讯作者：
Parrinello, M
影响因子：
16.6
作者：
Lira-Navarrete, Erandi;Iglesias-Fernandez, Javier;Hurtado-Guerrero, Ramon
通讯作者：
Hurtado-Guerrero, Ramon
影响因子：
6.8
作者：
Ficko-Blean, Elizabeth;Boraston, Alisdair B.
通讯作者：
Boraston, Alisdair B.
影响因子：
4.8
作者：
Kato, Kentaro;Jeanneau, Charlotte;Clausen, Henrik
通讯作者：
Clausen, Henrik