Wnt 同工型与共受体的特异性相互作用决定了 LRP6 抗体对信号的抑制或增强作用。

Wnt isoform-specific interactions with coreceptor specify inhibition or potentiation of signaling by LRP6 antibodies.

机构信息

Department of Cancer Targets, Genentech Research & Early Development, South San Francisco, California, United States of America.

出版信息

PLoS One. 2010 Sep 13;5(9):e12682. doi: 10.1371/journal.pone.0012682.

Abstract

β-Catenin-dependent Wnt signaling is initiated as Wnt binds to both the receptor FZD and coreceptor LRP5/6, which then assembles a multimeric complex at the cytoplasmic membrane face to recruit and inactivate the kinase GSK3. The large number and sequence diversity of Wnt isoforms suggest the possibility of domain-specific ligand-coreceptor interactions, and distinct binding sites on LRP6 for Wnt3a and Wnt9b have recently been identified in vitro. Whether mechanistically different interactions between Wnts and coreceptors might mediate signaling remains to be determined. It is also not clear whether coreceptor homodimerization induced extracellularly can activate Wnt signaling, as is the case for receptor tyrosine kinases. We generated monoclonal antibodies against LRP6 with the unexpected ability to inhibit signaling by some Wnt isoforms and potentiate signaling by other isoforms. In cell culture, two antibodies characterized further show reciprocal activities on most Wnts, with one antibody antagonizing and the other potentiating. We demonstrate that these antibodies bind to different regions of LRP6 protein, and inhibition of signaling results from blocking Wnt binding. Antibody-mediated dimerization of LRP6 can potentiate signaling only when a Wnt isoform is also able to bind the complex, presumably recruiting FZD. Endogenous autocrine Wnt signaling in different tumor cell lines can be either antagonized or enhanced by the LRP6 antibodies, indicating expression of different Wnt isoforms. As anticipated from the roles of Wnt signaling in cancer and bone development, antibody activities can also be observed in mice for inhibition of tumor growth and in organ culture for enhancement of bone mineral density. Collectively, our results indicate that separate binding sites for different subsets of Wnt isoforms determine the inhibition or potentiation of signaling conferred by LRP6 antibodies. This complexity of coreceptor-ligand interactions may allow for differential regulation of signaling by Wnt isoforms during development, and can be exploited with antibodies to differentially manipulate Wnt signaling in specific tissues or disease states.

摘要

β-连环蛋白依赖性 Wnt 信号转导是由 Wnt 与受体 FZD 和共受体 LRP5/6 结合引发的,随后在细胞质膜表面组装一个多聚体复合物,募集并失活激酶 GSK3。Wnt 同工型的数量和序列多样性表明存在特定配体-共受体相互作用的可能性,并且最近在体外鉴定了 LRP6 上 Wnt3a 和 Wnt9b 的不同结合位点。Wnt 和共受体之间的机制上不同的相互作用是否可能介导信号转导仍有待确定。也不清楚共受体的细胞外同源二聚化是否可以激活 Wnt 信号转导,就像受体酪氨酸激酶一样。我们生成了针对 LRP6 的单克隆抗体,具有意想不到的抑制某些 Wnt 同工型信号转导和增强其他同工型信号转导的能力。在细胞培养中,进一步表征的两种抗体对大多数 Wnt 表现出相互作用的活性,一种抗体拮抗,另一种抗体增强。我们证明这些抗体结合 LRP6 蛋白的不同区域,信号转导的抑制是由于阻断 Wnt 结合。只有当 Wnt 同工型也能够结合复合物时,LRP6 的抗体介导的二聚化才能增强信号转导,推测募集 FZD。不同肿瘤细胞系中的内源性自分泌 Wnt 信号转导可以被 LRP6 抗体拮抗或增强,表明表达不同的 Wnt 同工型。正如 Wnt 信号转导在癌症和骨骼发育中的作用所预期的那样,抗体活性也可以在抑制肿瘤生长的小鼠中和增强骨密度的器官培养中观察到。总之,我们的结果表明,不同 Wnt 同工型亚群的分离结合位点决定了 LRP6 抗体赋予的信号转导的抑制或增强。这种共受体-配体相互作用的复杂性可能允许 Wnt 同工型在发育过程中对信号转导进行差异调节,并且可以利用抗体在特定组织或疾病状态下差异地操纵 Wnt 信号转导。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ae80/2938341/876a0268c048/pone.0012682.g001.jpg

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