title: Convulxin, a C-type lectin-like protein, inhibits HCASMCs functions via WAD-motif/integrin-αv interaction and NF-κB-independent gene suppression of GRO and IL-8.
abstract: Convulxin (CVX), a C-type lectin-like protein (CLPs), is a potent platelet aggregation inducer. To evaluate its potential applications in angiogenic diseases, the multimeric CVX were further explored on its mode of actions toward human coronary artery smooth muscle cells (HCASMCs). The N-terminus of β-chain of CVX (CVX-β) contains a putative disintegrin-like domain with a conserved motif upon the sequence comparison with other CLPs. Importantly, native CVX had no cytotoxic activity as examined by electrophoretic pattern. A Trp-Ala-Asp (WAD)-containing octapeptide, MTWADAEK, was thereafter synthesized and analyzed in functional assays. In the case of specific integrin antagonists as positive controls, the anti-angiogenic effects of CVX on HCASMCs were investigated by series of functional analyses. CVX showed to exhibit multiple inhibitory activities toward HCASMCs proliferation, adhesion and invasion with a dose- and integrin αvβ3-dependent fashion. However, the WAD-octapeptide exerting a minor potency could also work as an active peptidomimetic. In addition, flow cytometric analysis demonstrated both the intact CVX and synthetic peptide can specifically interact with integrin-αv on HCASMCs and CVX was shown to have a down-regulatory effect on the gene expression of CXC-chemokines, such as growth-related oncogene and interleukin-8. According to nuclear factor-κB (NF-κB) p65 translocation assay and Western blotting analysis, the NF-κB activation was not involved in the signaling events of CVX-induced gene expression. In conclusion, CVX may act as a disintegrin-like protein via the interactions of WAD-motif in CVX-β with integrin-αv on HCASMCs and it also is a gene suppressor with the ability to diminish the expression of two CXC-chemokines in a NF-κB-independent manner. Indeed, more extensive investigations are needed and might create a new avenue for the development of a novel angiostatic agent.
authors.affiliation: Chang Gung University of Science and Technology, Guishan Dist., Taoyuan City, Taiwan.
authors.forename: Chun-Ho
authors.initials: CH
authors.lastname: Shih
authors.affiliation: Chang Gung University of Science and Technology, Guishan Dist., Taoyuan City, Taiwan.
authors.forename: Tin-Bin
authors.initials: TB
authors.lastname: Chiang
authors.affiliation: Chang Gung University of Science and Technology, Guishan Dist., Taoyuan City, Taiwan; Department of Neurological Surgery, Chang Gung Memorial Hospital, Guishan Dist., Taoyuan City, Taiwan. Electronic address: wjwang@mail.cgust.edu.tw.
authors.forename: Wen-Jeng
authors.initials: WJ
authors.lastname: Wang
mesh_terms: D001665:Binding Sites; D002448:Cell Adhesion; D002460:Cell Line; D049109:Cell Proliferation; D054360:Chemokine CXCL1 / Q000235:genetics / Q000378:metabolism*; D003331:Coronary Vessels / Q000166:cytology; D003435:Crotalid Venoms / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D006801:Humans; D039564:Integrin alphaV / Q000378:metabolism*; D016209:Interleukin-8 / Q000378:metabolism*; D037181:Lectins, C-Type / Q000737:chemistry / Q000378:metabolism*; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000378:metabolism; D032389:Myocytes, Smooth Muscle / Q000378:metabolism*; D016328:NF-kappa B / Q000378:metabolism; D011485:Protein Binding
chemical_list: C475829:CXCL1 protein, human; D054360:Chemokine CXCL1; D003435:Crotalid Venoms; D039564:Integrin alphaV; D016209:Interleukin-8; D037181:Lectins, C-Type; D016328:NF-kappa B; C028890:convulxin
keywords: CXC-chemokine; Convulxin; Disintegrin; HCASMC; Integrin-αv; WAD