Laminin Receptor Involvement in the Anti-angiogenic Activity of Pigment Epithelium-derived Factor
Laminin Receptor Involvement in the Anti-angiogenic Activity of Pigment Epithelium-derived Factor
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DOI：
10.1074/jbc.m809259200
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发表时间：
2009-04-17
影响因子：
4.8
通讯作者：
Li, Zhenlin
中科院分区：
文献类型：
--
作者：
Bernard, Adrien;Gao-Li, Jacqueline;Li, Zhenlin
色素上皮衍生因子(PEDF)是一种多功能蛋白质，具有神经营养、抗氧化和抗炎等特性。它也是最有效的内源性血管生成抑制因子之一，在抑制肿瘤生长、侵袭和转移方面发挥着重要作用。研究表明，PEDF可以与细胞表面蛋白结合，但人们对其如何发挥作用知之甚少。最近，研究发现磷脂酶A(2)/Nutrin/Patatin样磷脂酶结构域包含2是一种PEDF受体。为了识别其他受体，我们以PEDF为诱饵进行酵母双杂交筛选，发现非整合素37/67-kDa层粘连蛋白受体(LR)是另一种PEDF受体。免疫共沉淀、His标签下拉和表面等离子体共振分析证实了PEDF和LR之间的相互作用。利用酵母双杂交的方法，我们进一步将PEDF上的LR相互作用结构域限制为34个氨基酸(AA)多肽(a44-77)，将LR上的PEDF相互作用结构域限制为91-aa片段(aa120-210)。一种命名为P46(aa46-70)的25聚体多肽来源于34聚体，在表面等离子体共振分析中与LR相互作用，并结合到内皮细胞(EC)膜上。该肽可诱导EC凋亡，抑制EC迁移、体外管状网络形成和体外视网膜血管生成，与PEDF类似。我们的结果表明，LR是一种真正的PEDF受体，介导PEDF血管生成抑制。
Pigment epithelium-derived factor (PEDF) is a multifunctional protein with neurotrophic, anti-oxidative, and anti-inflammatory properties. It is also one of the most potent endogenous inhibitors of angiogenesis, playing an important role in restricting tumor growth, invasion, and metastasis. Studies show that PEDF binds to cell surface proteins, but little is known about how it exerts its effects. Recently, research identified phospholipase A(2)/nutrin/patatin-like phospholipase domain-containing 2 as one PEDF receptor. To identify other receptors, we performed yeast two-hybrid screening using PEDF as bait and discovered that the non-integrin 37/67-kDa laminin receptor (LR) is another PEDF receptor. Co-immunoprecipitation, His tag pulldown, and surface plasmon resonance assays confirmed the interaction between PEDF and LR. Using the yeast two-hybrid method, we further restricted the LR-interacting domain on PEDF to a 34-amino acid (aa) peptide (aa 44-77) and the PEDF-interacting domain on LR to a 91-aa fragment (aa 120-210). A 25-mer peptide named P46 (aa 46-70), derived from 34-mer, interacts with LR in surface plasmon resonance assays and binds to endothelial cell (EC) membranes. This peptide induces EC apoptosis and inhibits EC migration, tube-like network formation in vitro, and retinal angiogenesis ex vivo, like PEDF. Our results suggest that LR is a real PEDF receptor that mediates PEDF angiogenesis inhibition.