The role of proteases in regulating Eph/ephrin signaling

Proteases regulate a myriad of cell functions, both in normal and disease states. In addition to protein turnover, they regulate a range of signaling processes, including those mediated by Eph receptors and their ephrin ligands. A variety of proteases is reported to directly cleave Ephs and/or ephrins under different conditions, to promote receptor and/or ligand shedding, and regulate receptor/ligand internalisation and signaling. They also cleave other adhesion proteins in response to Eph-ephrin interactions, to indirectly facilitate Eph-mediated functions. Proteases thus contribute to Eph/ephrin mediated changes in cell-cell and cell-matrix interactions, in cell morphology and in cell migration and invasion, in a manner which appears to be tightly regulated by, and co-ordinated with, Eph signaling. This review summarizes the current literature describing the function and regulation of protease activities during Eph/ephrin-mediated cell signaling.     

Eph receptor and ephrin function in breast,gut, and skin epithelia

Epithelial cells are tightly coupled together through specialized intercellular junctions, including adherens junctions, desmosomes, tight junctions, and gap junctions. A growing body of evidence suggests epithelial cells also directly exchange information at cell-cell contacts via the Eph family of receptor tyrosine kinases and their membrane-associated ephrin ligands. Ligand-dependent and -independent signaling via Eph receptors as well as reverse signaling through ephrins impact epithelial tissue homeostasis by organizing stem cell compartments and regulating cell proliferation, migration, adhesion, differentiation, and survival. This review focuses on breast, gut, and skin epithelia as representative examples for how Eph receptors and ephrins modulate diverse epithelial cell responses in a context-dependent manner. Abnormal Eph receptor and ephrin signaling is implicated in a variety of epithelial diseases raising the intriguing possibility that this cell-cell communication pathway can be therapeutically harnessed to normalize epithelial function in pathological settings like cancer or chronic inflammation.     

To phosphorylate or not to phosphorylate: Selective alterations in tyrosine kinase-inhibited EphB mutant mice

EphB tyrosine kinase receptors have been implicated in multiple developmental processes; however, the signaling mechanism underlying these events remains unclear. Through a triple knock-in mouse line for three neurally expressed EphBs, Sokis et al. demonstrated that EphB tyrosine kinase activity is required for axon guidance but does not influence synapse formation. This short communication highlights their study and appealing molecular approach that elucidated the functions of EphB tyrosine kinase during developmental events.     

Crystal structure of the ligand-binding domain of the promiscuous EphA4 receptor reveals two distinct conformations

Eph receptors and their ephrin ligands are important mediators of cell-cell communication. They are divided in two subclasses based on their affinities for each other and on sequence conservation. Receptor-ligand binding within each subclass is fairly promiscuous, while binding cross the subclasses happens rarely. EphA4 is an exception to this general rule, since it has long been known to bind both A- and B-class ephrin ligands but the reason for this exceptional behavior has not been worked out at molecular level. Recent structural and biochemical studies on EphA4 ligand-binding domain alone and in complex with its ligands have addressed this question. However, the published structures of EphA4/ephrin complexes differ considerably from each other and strikingly different explanations for the exceptional promiscuity of EphA4 were proposed. To address these contradictory findings, we have determined a crystal structure of the EphA4 ligand-binding domain at 2.3 Å resolution and show that the receptor has an unprecedented ability to exist in two very different, well-ordered conformations even in the unbound state. Our results suggest that the ligand promiscuity of the Ephs is directly correlated with the structural flexibility of the ligand-binding surface of the receptor.     

Eph-ephrin介导反向信号传递的研究进展

双向信号传递是细胞间通讯领域中新近阐明的机制,酪氨酸激酶受体-配体(Eph-ephrin)介导的双向信号传递是此机制中的一个重要代表.Eph酪氨酸激酶家族受体及其配体ephrin家族成员是在神经发育、血管新生等方面起重要作用的分子,通过Eph向细胞内传递的信号称为正向信号,通过其配体ephrin的信号称为反向信号.Ephrin家族又可根据分子结构分为2个亚家族,其中ephrinB为跨膜蛋白,可通过酪氨酸磷酸化依赖和PDZ结合结构域介导2种方式向胞内传递反向信号,活化FAK、JNK、Wnt等信号通路,ephrinA为糖基磷脂酰肌醇锚定蛋白,也具有反向信号传递功能.     

Ephrin-B2 selectively marks arterial vessels and neovascularization sites in the adult, with expression in both endothelial and smooth-muscle cells

The Eph receptor tyrosine kinases and their membrane-tethered ephrin ligands provide critical guidance cues at points of cell-to-cell contact. It has recently been reported that the ephrin-B2 ligand is a molecular marker for the arterial endothelium at the earliest stages of embryonic angiogenesis, while its receptor EphB4 reciprocally marks the venous endothelium. These findings suggested that ephrin-B2 and EphB4 are involved in establishing arterial versus venous identity and perhaps in anastamosing arterial and venous vessels at their junctions. By using a genetically engineered mouse in which the lacZ coding region substitutes and reports for the ephrin-B2 coding region, we demonstrate that ephrin-B2 expression continues to selectively mark arteries during later embryonic development as well as in the adult. However, as development proceeds, we find that ephrin-B2 expression progressively extends from the arterial endothelium to surrounding smooth muscle cells and to pericytes, suggesting that ephrin-B2 may play an important role during formation of the arterial muscle wall. Furthermore, although ephrin-B2 expression patterns vary in different vascular beds, it can extend into capillaries about midway between terminal arterioles and postcapillary venules, challenging the classical conception that capillaries have neither arterial nor venous identity. In adult settings of angiogenesis, as in tumors or in the female reproductive system, the endothelium of a subset of new vessels strongly expresses ephrin-B2, once again contrary to earlier views that such new vessels lack arterial/venous characteristics and derive from postcapillary venules. While earlier studies had focused on a role for ephrin-B2 during the earliest embryonic stages of arterial/venous determination, our current findings using ephrin-B2 as an arterial marker in the adult challenge prevailing views of the arterial/venous identity of quiescent as well as remodeling adult microvessels and also highlight a possible role for ephrin-B2 in the formation of the arterial muscle wall.     

促红细胞生成素产生肝细胞受体A2(EphA2)SAM结构域对激酶结构域的脂质体结合能力与激酶活性的调控研究

促红细胞生成素产生肝细胞受体(Eph receptor) 是受体酪氨酸激酶(RTK)家族中最大的亚家族,其介导的双向信号传导对细胞的形态、黏附、运动、增殖、生存及分化都有重要的调控作用。EphA2是Eph受体家族中一个被广泛研究的重要亚型,在白内障和乳腺癌等病理发生过程中发挥了重要作用。既往研究发现:EphA2受体的激酶结构域可结合细胞膜,其激酶活性受磷脂膜的调控,但是相邻的SAM结构域对激酶结构域与脂膜的相互作用以及激酶活性的影响尚不清楚。在此项研究中,通过与磷酸酶PTP1B1-301活性片段共表达的方式,表达、纯化了EphA2受体的胞内段激酶-SAM串联结构域,通过比较胞内段激酶-SAM串联结构域与单独激酶结构域的脂质体结合能力,以及测定对应的激酶活性,发现:EphA2受体胞内段的SAM结构域使其激酶结构域与脂质体(4 mg/mL)的结合能力增强约6倍(P＜0.001);磷酸化后的EphA2胞内段激酶-SAM串联结构域结合脂质体(4 mg/mL)的能力比非磷酸化的胞内段激酶-SAM串联结构域提高2.5倍(P＜0.05);而结合脂质体后,激酶结构域的激酶活性也被进一步提高,从而形成正反馈。综上所述,本研究的发现提示:EphA2胞内段的酪氨酸激酶结构域与相邻的SAM结构域可形成一个完整的结构功能单位,其激酶活性和脂质体结合能力与单独的激酶结构域相比都形成了明显的差异,我们的这一发现对进一步理解Eph受体家族其他亚型的激酶结构域的活性调控提供了参考与思路。     

Regulation of signaling interactions and receptor endocytosis in growing blood vessels

Blood vessels and the lymphatic vasculature are extensive tubular networks formed by endothelial cells that have several indispensable functions in the developing and adult organism. During growth and tissue regeneration but also in many pathological settings, these vascular networks expand, which is critically controlled by the receptor EphB4 and the ligand ephrin-B2. An increasing body of evidence links Eph/ephrin molecules to the function of other receptor tyrosine kinases and cell surface receptors. In the endothelium, ephrin-B2 is required for clathrin-dependent internalization and full signaling activity of VEGFR2, the main receptor for vascular endothelial growth factor. In vascular smooth muscle cells, ephrin-B2 antagonizes clathrin-dependent endocytosis of PDGFRβ and controls the balanced activation of different signal transduction processes after stimulation with platelet-derived growth factor. This review summarizes the important roles of Eph/ephrin molecules in vascular morphogenesis and explains the function of ephrin-B2 as a molecular hub for receptor endocytosis in the vasculature.     

Eph受体家族及其配体的信号转导途径及功能

Eph受体是已知最大的酪氨酸蛋白激酶受体家族,Eph受体和其膜附着型配体（ephrin）在发育过程中呈现不同的表达模式,近来研究证明,Eph受体和其配体在包括神经网络形成,神经管和轴旁中胚层的成型（patterning）,细胞迁移导向和轴突路径导引,血管形成等许多的发育过程中起重要作用.Eph受体及其配体也与肿瘤发生有关,因此深入分析这些分子尤其在肿瘤细胞生长中的功能而应用于治疗具有重要的临床意义.     

Reelin induces EphB activation

The integration of newborn neurons into functional neuronal networks requires migration of cells to their final position in the developing brain, the growth and arborization of neuronal processes and the formation of synaptic contacts with other neurons. A central player among the signals that coordinate this complex sequence of differentiation events is the secreted glycoprotein Reelin, which also modulates synaptic plasticity, learning and memory formation in the adult brain. Binding of Reelin to ApoER2 and VLDL receptor, two members of the LDL receptor family, initiates a signaling cascade involving tyrosine phosphorylation of the intracellular cytoplasmic adaptor protein Disabled-1, which targets the neuronal cytoskeleton and ultimately controls the positioning of neurons throughout the developing brain. However, it is possible that Reelin signals interact with other receptor-mediated signaling cascades to regulate different aspects of brain development and plasticity. EphB tyrosine kinases regulate cell adhesion and repulsion-dependent processes via bidirectional signaling through ephrin B transmembrane proteins. Here, we demonstrate that Reelin binds to the extracellular domains of EphB transmembrane proteins, inducing receptor clustering and activation of EphB forward signaling in neurons, independently of the ''classical'' Reelin receptors, ApoER2 and VLDLR. Accordingly, mice lacking EphB1 and EphB2 display a positioning defect of CA3 hippocampal pyramidal neurons, similar to that in Reelin-deficient mice, and this cell migration defect depends on the kinase activity of EphB proteins. Together, our data provide biochemical and functional evidence for signal integration between Reelin and EphB forward signaling.