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The Structure and Functions of Paxillin and Its Roles in Neovascularization

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The Structure and Functions of Paxillin and Its Roles in Neovascularization European Review for Medical and Pharmacological Sciences 2017; 21: 1768-1773 The structure and functions of paxillin and its roles in neovascularization W.-J. YANG, J. ZHONG, J.-G. YU, F. ZHAO, Y. XIANG 1Department of Ophthalmology, Key Laboratory for Molecular Diagnosis of Hubei Province, the Central Hospital of Wuhan, Tongji Medical College of Huazhong University of Science and Technology, Wuhan, China Wanju Yang and Jie Zhong contributed equally to this work Abstract. – Identifying and studying the mo- tion, which leads to the formation of tight connec- lecular mechanisms of neovascularization bio- tions between lamellar collagen structures. This markers are critical for conquering many diseas- anatomical structure results in failed migration es, such as corneal diseases and cancer. Paxil- of vascular endothelial cells in the cornea1. The lin is an important cell scaffold and cellular sig- naling protein, especially a key molecule of the corneal avascular state is the key to maintaining Integrin-mediated downstream signaling trans- corneal transparency and ensure good eyesight. duction. This review summarizes the structure However, a variety of corneal diseases, such as and functions of paxillin, and the research prog- infection, trauma, and chemical injury, may lead ress of its roles in neovascularization. Although to corneal neovascularization, which will damage there are still some problems to be solved, paxil- the transparency and normal microenvironment lin may become an important target of anti-neo- of the cornea, and cause the decrease of eye- vascularization therapies. sight and even blindness2,3. Meanwhile, corneal Key Words: neovascularization makes the intrinsic immune Paxillin, Neovascularization, Integrin, Vascular en- privilege disappear and is a high-risk factor of dothelial growth factor. immunologic rejection after corneal transplanta- tion. Currently, 4.14% of the ophthalmic patients in America suffer from corneal neovasculariza- Abbreviations tion4. The corneal neovascularization problem caused by various eye diseases, and eventually LD: lethal dose; LIM: limit; ERK: extracellular sig- leading to blindness, has become one of most nal-regulated kinas; MEK: Methyl Ethyl Ketone; ARF: serious ophthalmic problems5. However, it is still acute renal failure; PKL: paxillin kinase linker; PIX: lack of efficient and simple method to treat cor- Proton induced X-ray; PAK: p21-activated kinase; FAK: focal adhesion kinase; MDCK: Madin-Darby canine neal neovascularization. The key biological char- kidney; EPCs: endothelial progenitor cells; FLK-1: Fe- acteristics of neovascularization are infiltration tal liver kinase-1; VEGF: vascular endothelial growth and metastasis. With the progress of cellular and factor. molecular biology, many recent kinds of research have focused on studying the molecular mecha- nisms of corneal neovascularization and found that gene regulation is closely related to corneal Introduction neovascularization. Inactivation, mutation, and overexpression of certain genes play important roles in corneal neovascularization. Therefore, With the research progress on the molecu- identifying and studying neovascularization-re- lar mechanisms of angiogenesis, molecular tar- lated biomarkers are critical for conquering the get-based treatment may bring a broad prospect problem of neovascularization. Paxillin is a mul- to the clinic treatment of neovascularization. Nor- tidomain protein discovered in recent years. It is mally, the cornea is in an avascular state, which an important cell scaffold and cellular signaling is called “angiogenic privilege.” An important protein, especially a key molecule of the Integ- cause of this state is continuous corneal dehydra- rin-mediated downstream signaling transduction. 1768 Corresponding Author: Yi Xiang, MD; e-mail: [email protected] The structure and functions of paxillin and its roles in neovascularization Through protein-protein interactions and under site tyrosine 118 can induce the change of pax- the regulation by phosphorylation, paxillin ex- illin structure and lead to the binding of ERK, pression shows dynamic changes, which affects which causes the accumulation of a large amount focal adhesion, cell proliferation, adhesion, mi- of ERK proteins at the focal adhesion12. Also, gration, survival, dissemination, and cytoskele- paxillin can bind to Raf and MEK at the focal ton reconstruction. The metastasis and infiltra- adhesion to activate the ERK pathway, which can tion of new vessels are closely related to the above enhance cell diffusion and mobility13. After the cellular processes. It is presumed that paxillin binding of ARF-GAP/PKL complex with paxil- can also regulate the development and metastatic lin LD4 domain, PIX and PAK are successively potential of new vessels. This review summarizes activated to trigger Cdc42/Rac activities at the the structure and functions of paxillin and the re- focal adhesion, which can inhibit Rho expression search progress of its roles in neovascularization. and promote directional cell and lamellipodia ex- tension14,15. Paxillin contains multiple phosphory- Chemical Structure of Paxillin lation sites and structural domains and, thus, can Paxillin is a phosphoprotein with the molecular bind to the above signal proteins and structural weight of 68 kD6. It was firstly discovered in the proteins, which makes it a “transfer station” of fibroblasts transformed by Rous sarcoma virus signaling transduction. in 1989. Paxillin is primarily located in local focal adhesion and it is an important component LD Domain of the focal adhesion. It participates in focal LD domain is located at the N-terminal of adhesion assembly and can bind to focal adhe- paxillin and regulates the signal transduction of sion proteins and actin. Paxillin has 11 exomes paxillin. Its binding site to FAK and Vinculin and encodes 559 aminoacids. Human paxillin is is highly conserved16, with five leucine and as- located on chromosome 12q24. The N-terminal partic acid-binding motifs (LD1-LD5) and the of paxillin consists of 5 LD motifs, which are common sequence LDXLLXXL. It is considered involved in protein recognition. The C-terminal as the binding platform for FAK and Vinculin7. of paxillin consists of four tandem LIM domains, Although these motifs are similar, they have dif- which mediate protein-protein interactions. Also, ferent protein-protein interactions. LD1 mediates its N-terminal contains several proline-rich SH3 the interactions to Actopaxin17 (it is located at the binding motifs. Also, it has many serine/thre- focal adhesion and serves as the binding protein onine and tyrosine phosphorylation sites, which of paxillin and Actin), Integrin-linked kinase are important intermediate components of cell (ILK), Vinculin and HPV E6 protein. LD2 binds signaling transduction. The multiple domains of to FAK, Vinculin and the proline-rich Tyrosine paxillin can bind to a series of signaling proteins kinase 2. LD4 binds to Actopaxin18. LD3 has and structural proteins, and mediate cell-signal- been degenerated and lacks the conserved se- ing transduction. It has been found that paxillin quence. Paxillin kinase linker may bind to focal plays important roles in cell adhesion and mi- adhesion kinase 3, Clathrin and polyA-binding gration. Previous studies7 showed that paxillin protein19. So far, although the phosphorylation can serve as a downstream signal of the FAK/ of FAK tyrosine in embryonic stem cells needs Src signaling pathway and be phosphorylated. LD5, the proteins that can bind to the degenerated The paxillin phosphorylation sites at tyrosines 31 LD3 domain have not been found yet20. Paxillin and 118 can bind to Crk SH2 domain, which pro- LD4 domain is a particularly important site for motes the formation of Crk-DOCK180 complex regulating Rho GTPase signals. and, thus, activate the Rac pathway to enhance lamellipodia extension and increase cell mobility. LIM Domain Also, Crk SH3 domain can bind to C3G (a Ras The C-terminal of paxillin has four tandem guanine nucleotide exchanging factor) to activate domains, and each domain consists of a double Ras pathway8,9. Paxillin tyrosines 31 and 118 can zinc-finger motif and includes two cysteine- and also bind to the SH2 domain of p120 Ras GAP histidine-rich sequences. This special structure to separate p190 RhoGAP from itself. Free p190 is initially found in the homeodomain proteins RhoGAP can inhibit RhoA in the focal adhesion Lin-11, Isl-1 and Mec-3. Therefore, it is called and, thus, promote Rac-mediated lamellipodia the LIM domain21. The two zinc finger structures extension and enhance cell migration10,11. The of LIM domain are bundled together by hydro- binding of Src and the paxillin phosphorylation phobic interaction. Every zinc finger consists of 1769 W.-J. Yang, J. Zhong, J.-G. Yu, F. Zhao, Y. Xiang two antiparallel β-sheets, which are separated by cell adhesion and mobility in culture medium are a turn and have a short α-helix at the terminal. decreased23. Paxillin-deficient fibroblasts can im- The third LIM domain is the critical structure pair cell migration, which indicates that paxillin for paxillin to bind with focal adhesion proteins. can regulate cell dissemination and mobility24. LIM domain not only mediates the localization of The assembly and disassembly of focal adhe- paxillin on the focal adhesion and actin cytoskel- sion regulate cell adhesion and movement, thus etons, but also mediates the interactions to many affecting tumor metastasis. These processes are proteins including PTP-PEST,
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