Crystallization of Galectin-8 Linker Reveals Intricate Relationship between the N-terminal Tail and the Linker.
Crystallization of Galectin-8 Linker Reveals Intricate Relationship between the N-terminal Tail and the Linker.
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DOI：
10.3390/ijms17122088
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发表时间：
2016-12-12
影响因子：
5.6
通讯作者：
Su J
中科院分区：
文献类型：
--
作者：
Si Y;Wang Y;Gao J;Song C;Feng S;Zhou Y;Tai G;Su J
半乳糖凝集素-8(Galectin-8，Gal-8)在正常免疫功能和癌症中发挥着重要作用。该凝集素含有两个由多肽接头连接的碳水化合物识别结构域(CRD)。N-末端CRD决定了配体结合的特异性，而连接子被认为调节Gal-8的整体功能，包括多聚体和生物活性。在这里，我们使用含有Ni2+的结晶条件，将Gal-8 N-末端CRD与多肽连接物结晶。Ni2+离子在两个CRD之间通过晶体堆积接触形成络合。Ni2+与Asp25之间的配位作用在决定β链F0的结构和影响连接子构象方面起着间接作用，但由于其动力学性质而无法确定。连接体还被原位缩短化，并在不同的条件下结晶，导致了更高的分辨率结构，细化到1.08ä。这种晶体结构允许定义一小部分连接体通过离子相互作用和氢键与Gal-8 N末端尾部相互作用。对两个Gal-8 N-末端CRD结构的观察表明，N-末端的尾部和连接子可能会相互影响彼此的构象。此外，在特定的结晶条件下，甘油可以取代乳糖，并在碳水化合物结合部位观察到。然而，甘油在血凝试验中没有显示抑制活性。
Galectin-8 (Gal-8) plays a significant role in normal immunological function as well as in cancer. This lectin contains two carbohydrate recognition domains (CRD) connected by a peptide linker. The N-terminal CRD determines ligand binding specificity, whereas the linker has been proposed to regulate overall Gal-8 function, including multimerization and biological activity. Here, we crystallized the Gal-8 N-terminal CRD with the peptide linker using a crystallization condition that contains Ni2+. The Ni2+ ion was found to be complexed between two CRDs via crystal packing contacts. The coordination between Ni2+ and Asp25 plays an indirect role in determining the structure of β-strand F0 and in influencing the linker conformation which could not be defined due to its dynamic nature. The linker was also shortened in situ and crystallized under a different condition, leading to a higher resolution structure refined to 1.08 Å. This crystal structure allowed definition of a short portion of the linker interacting with the Gal-8 N-terminal tail via ionic interactions and hydrogen bonds. Observation of two Gal-8 N-terminal CRD structures implies that the N-terminal tail and the linker may influence each other’s conformation. In addition, under specific crystallization conditions, glycerol could replace lactose and was observed at the carbohydrate binding site. However, glycerol did not show inhibition activity in hemagglutination assay.
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影响因子：
4.8
作者：
HADARI, YR;PAZ, K;ZICK, Y
通讯作者：
ZICK, Y
影响因子：
3
作者：
Bidon-Wagner, N;Le Pennec, JP
通讯作者：
Le Pennec, JP
DOI：
10.1107/s0907444909052925
发表时间：
2010-02
期刊：
Acta crystallographica. Section D, Biological crystallography
影响因子：
--
作者：
Adams PD;Afonine PV;Bunkóczi G;Chen VB;Davis IW;Echols N;Headd JJ;Hung LW;Kapral GJ;Grosse-Kunstleve RW;McCoy AJ;Moriarty NW;Oeffner R;Read RJ;Richardson DC;Richardson JS;Terwilliger TC;Zwart PH
通讯作者：
Zwart PH
影响因子：
14.9
作者：
Davis IW;Leaver-Fay A;Chen VB;Block JN;Kapral GJ;Wang X;Murray LW;Arendall WB 3rd;Snoeyink J;Richardson JS;Richardson DC
通讯作者：
Richardson DC
DOI：
10.1107/s0907444909042073
发表时间：
2010-01
期刊：
Acta crystallographica. Section D, Biological crystallography
影响因子：
--
作者：
Chen VB;Arendall WB 3rd;Headd JJ;Keedy DA;Immormino RM;Kapral GJ;Murray LW;Richardson JS;Richardson DC
通讯作者：
Richardson DC