Monomerization of viral entry inhibitor griffithsin elucidates the relationship between multivalent binding to carbohydrates and anti-HIV activity.
Monomerization of viral entry inhibitor griffithsin elucidates the relationship between multivalent binding to carbohydrates and anti-HIV activity.
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DOI：
10.1016/j.str.2010.05.016
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发表时间：
2010-09-08
期刊：
影响因子：
--
通讯作者：
Wlodawer A
中科院分区：
文献类型：
--
作者：
Moulaei T;Shenoy SR;Giomarelli B;Thomas C;McMahon JB;Dauter Z;O'Keefe BR;Wlodawer A
将突变引入抗病毒凝集素griffithsin（GRFT）的结构域交换的同源二聚体。而几个单和双突变体保持二聚体，在二聚化界面插入两个或四个氨基酸导致蛋白质的单体形式（mGRFT）。修饰的蛋白质的单体特征通过沉降平衡超离心和它们的高分辨率X-射线晶体结构来证实，而它们与碳水化合物的结合通过等温滴定量热法来评估。利用不同mGRFT变体的基于细胞的抗病毒活性测定表明，凝集素的单体形式对HIV-1的活性大大降低，表明GRFT的抗病毒活性源于病毒颗粒通过GRFT和HIV包膜糖蛋白上存在的寡糖之间的多价相互作用而交联和聚集。mGRFT和nonamannoside之间的复合物的原子分辨率晶体结构显示，单个mGRFT分子通过单体上存在的所有三个碳水化合物结合位点与两个不同的nonamannoside分子结合。<$天然griffithsin是一种结构域交换的二聚体，具有很强的抗病毒活性<$通过结构导向设计获得了griffithsin的几种单体突变体<$HIV gp 120相关的寡糖被证明可以交联griffithsin分子<$二聚体结构和交联是维持抗病毒活性所必需的
Mutations were introduced to the domain-swapped homodimer of the antiviral lectin griffithsin (GRFT). Whereas several single and double mutants remained dimeric, insertion of either two or four amino acids at the dimerization interface resulted in a monomeric form of the protein (mGRFT). Monomeric character of the modified proteins was confirmed by sedimentation equilibrium ultracentrifugation and by their high resolution X-ray crystal structures, whereas their binding to carbohydrates was assessed by isothermal titration calorimetry. Cell-based antiviral activity assays utilizing different variants of mGRFT indicated that the monomeric form of the lectin had greatly reduced activity against HIV-1, suggesting that the antiviral activity of GRFT stems from crosslinking and aggregation of viral particles via multivalent interactions between GRFT and oligosaccharides present on HIV envelope glycoproteins. Atomic resolution crystal structure of a complex between mGRFT and nonamannoside revealed that a single mGRFT molecule binds to two different nonamannoside molecules through all three carbohydrate-binding sites present on the monomer. ► Native griffithsin is a domain-swapped dimer with potent antiviral properties ► Several monomeric mutants of griffithsin were obtained by structure-guided design ► A HIV gp120-associated oligosaccharide was shown to crosslink molecules of griffithsin ► Dimeric structure and crosslinking are necessary for maintaining antiviral activity
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影响因子：
4.1
作者：
Bourne, Y;Astoul, CH;Rougé, P
通讯作者：
Rougé, P
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：
3.4
作者：
Schuck, P
通讯作者：
Schuck, P
影响因子：
5.7
作者：
Lubkowski, J;Hennecke, F;Wlodawer, A
通讯作者：
Wlodawer, A
影响因子：
2.9
作者：
PARKS, TD;LEUTHER, KK;DOUGHERTY, WG
通讯作者：
DOUGHERTY, WG