Molecular analysis of the pathophysiological binding of the platelet aggregation-inducing factor podoplanin to the C-type lectin-like receptor CLEC-2
Molecular analysis of the pathophysiological binding of the platelet aggregation-inducing factor podoplanin to the C-type lectin-like receptor CLEC-2
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DOI：
10.1111/j.1349-7006.2007.00634.x
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发表时间：
2008-01-01
期刊：
影响因子：
5.7
通讯作者：
Narimatsu, Hisashi
中科院分区：
文献类型：
--
作者：
Kato, Yukinari;Kaneko, Mika Kato;Narimatsu, Hisashi
粘蛋白型唾液酸糖蛋白podoplanin（aggrus）参与肿瘤细胞诱导的血小板聚集和肿瘤转移。C型凝集素样受体-2（CLEC-2）是血小板上podoplanin的内源性受体。然而，CLEC-2的病理生理学重要性和功能尚未阐明。在此，我们阐明了podoplanin和CLEC-2在体外和体内的病理生理相互作用。使用几个缺失突变体的CLEC-2表达为Fc嵌合体，我们首先确定了一个重要的podoplanin识别结构域的CLEC-2。此外，podoplanin-CLEC-2相互作用使用表达为Fc嵌合体的podoplanin的几种缺失突变体来证实。不仅二唾液酸-核心1-连接的糖肽，而且podoplanin蛋白的立体结构被发现是至关重要的CLEC-2结合活性的podoplanin。接下来，我们合成了podoplanin的各种糖肽，包括血小板聚集刺激结构域和Thr52上的O-聚糖。有趣的是，二唾液酸核心1连接的糖肽被CLEC-2特异性识别。此外，抗podoplanin单克隆抗体NZ-1抑制podoplanin-CLEC-2相互作用和podoplanin诱导的肺转移，表明CLEC-2是第一个被鉴定的podoplanin的病理生理受体。总之，我们阐明了血小板聚集诱导因子podoplanin及其在血小板上的特异性病理生理受体CLEC-2之间的体外和体内分子相互作用。Podoplanin和CLEC-2可能是癌症转移的有希望的治疗靶点。
The mucin-type sialoglycoprotein podoplanin (aggrus) is involved in tumor cell-induced platelet aggregation and tumor metastasis. C-type lectin-like receptor-2 (CLEC-2) was recently identified as an endogenous receptor of podoplanin on platelets. However, the pathophysiological importance and function of CLEC-2 have not been elucidated. Here we clarified the pathophysiological interaction between podoplanin and CLEC-2 in vitro and in vivo. Using several deletion mutants of CLEC-2 expressed as Fc chimeras, we first identified an important podoplanin-recognition domain in CLEC-2. Furthermore, the podoplanin-CLEC-2 interaction was confirmed using several deletion mutants of podoplanin expressed as Fc chimeras. Not only the disialyl-core1-attached glycopeptide but also the stereostructure of the podoplanin protein was found to be critical for the CLEC-2-binding activity of podoplanin. We next synthesized various glycopeptides of podoplanin that included both the platelet aggregation-stimulating domain and O-glycan on Thr52. Interestingly, a disialyl-core1-attached glycopeptide was recognized specifically by CLEC-2. Moreover, the anti-podoplanin monoclonal antibody NZ-1 suppressed both the podoplanin-CLEC-2 interaction and podoplanin-induced pulmonary metastasis, suggesting that CLEC-2 is the first pathophysiological receptor of podoplanin to be identified. In summary, we clarified the molecular interaction in vitro and in vivo between a platelet aggregation-inducing factor, podoplanin, and its specific pathophysiological receptor on platelets, CLEC-2. Podoplanin and CLEC-2 might represent promising therapeutic targets in cancer metastasis.