Modulation of ocular surface glycocalyx barrier function by a galectin-3 N-terminal deletion mutant and membrane-anchored synthetic glycopolymers.
Modulation of ocular surface glycocalyx barrier function by a galectin-3 N-terminal deletion mutant and membrane-anchored synthetic glycopolymers.
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DOI：
10.1371/journal.pone.0072304
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Argüeso P
中科院分区：
文献类型：
--
作者：
Mauris J;Mantelli F;Woodward AM;Cao Z;Bertozzi CR;Panjwani N;Godula K;Argüeso P
跨膜粘蛋白与多价碳水化合物结合蛋白半乳糖凝集素 3 的相互作用对于维持眼表面上皮糖萼的完整性至关重要。本研究旨在确定是否可以通过破坏半乳糖凝集素 3 多聚化和在质膜中插入合成糖聚合物来调节角膜上皮细胞的糖萼屏障功能。使用 siRNA 消除人角膜上皮细胞多层培养物中的半乳糖凝集素 3 生物合成以及半乳糖凝集素 3 无效小鼠中的半乳糖凝集素 3 生物合成，导致角膜屏障功能显着丧失，如对玫瑰红诊断染料的渗透性增加所表明的。将 β-乳糖（一种半乳糖凝集素 3 结合活性的竞争性碳水化合物抑制剂）添加到细胞培养系统中，会暂时破坏屏障功能。在这些实验中，使用缺乏 N 末端结构域的半乳糖凝集素 3 聚合显性负性抑制剂（而非全长半乳糖凝集素 3）进行治疗，可阻止屏障功能恢复至基础水平。通过荧光显微镜测定，含纤维二糖和含乳糖的糖聚合物均掺入角膜上皮细胞的顶膜中，与密集糖基化聚合物主链的链长分布无关。与含有乳糖的对应物相反，纤维二糖糖聚合物的膜掺入会损害角膜上皮细胞的屏障功能，后者在下拉测定中与半乳糖凝集素 3 结合。这些结果表明，半乳糖凝集素-3 多聚化和乳糖基残基的表面识别对于维持眼表面的糖萼屏障功能是必需的。半乳糖凝集素 3 结合的瞬时修饰可在治疗上用于提高局部药物递送的效率。
Interaction of transmembrane mucins with the multivalent carbohydrate-binding protein galectin-3 is critical to maintaining the integrity of the ocular surface epithelial glycocalyx. This study aimed to determine whether disruption of galectin-3 multimerization and insertion of synthetic glycopolymers in the plasma membrane could be used to modulate glycocalyx barrier function in corneal epithelial cells. Abrogation of galectin-3 biosynthesis in multilayered cultures of human corneal epithelial cells using siRNA, and in galectin-3 null mice, resulted in significant loss of corneal barrier function, as indicated by increased permeability to the rose bengal diagnostic dye. Addition of β-lactose, a competitive carbohydrate inhibitor of galectin-3 binding activity, to the cell culture system, transiently disrupted barrier function. In these experiments, treatment with a dominant negative inhibitor of galectin-3 polymerization lacking the N-terminal domain, but not full-length galectin-3, prevented the recovery of barrier function to basal levels. As determined by fluorescence microscopy, both cellobiose- and lactose-containing glycopolymers incorporated into apical membranes of corneal epithelial cells, independently of the chain length distribution of the densely glycosylated, polymeric backbones. Membrane incorporation of cellobiose glycopolymers impaired barrier function in corneal epithelial cells, contrary to their lactose-containing counterparts, which bound to galectin-3 in pull-down assays. These results indicate that galectin-3 multimerization and surface recognition of lactosyl residues is required to maintain glycocalyx barrier function at the ocular surface. Transient modification of galectin-3 binding could be therapeutically used to enhance the efficiency of topical drug delivery.
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影响因子：
15
作者：
Belardi, Brian;O'Donoghue, Geoff P.;Smith, Adam W.;Groves, Jay T.;Bertozzi, Carolyn R.
通讯作者：
Bertozzi, Carolyn R.
DOI：
10.1097/aci.0b013e32830e6b04
发表时间：
2008-10
影响因子：
2.8
作者：
Mantelli F;Argüeso P
通讯作者：
Argüeso P
影响因子：
15
作者：
Godula, Kamil;Bertozzi, Carolyn R.
通讯作者：
Bertozzi, Carolyn R.
影响因子：
4.2
作者：
Kompella UB;Kadam RS;Lee VH
通讯作者：
Lee VH
影响因子：
4.8
作者：
Earl, Lesley A.;Bi, Shuguang;Baum, Linda G.
通讯作者：
Baum, Linda G.