title: Voltage-Dependent Anion Channel-1, a Possible Ligand of Plasminogen Kringle 5.
abstract: Kringle 5, the fifth fragment of plasminogen, is known to be important for inhibiting the proliferation and migration of vascular endothelial cell (VEC), while not having any effects on normal endothelial cells. Therefore, it may be a potential tumor therapy candidate. However, the ligand of the Kringle 5 in VEC has not yet been identified. In this study, the possible ligand of Kringle 5 in vitro was screened and validated using Ph.D.-7 phage display peptide library with molecular docking, along with surface plasma resonance (SPR). After four rounds of panning, the specific clones of Kringle 5 were confirmed using enzyme-linked immunosorbent assay (ELISA). The gene sequence analysis showed that they expressed the common amino sequence IGNSNTL. Then, using a NCBI BLAST, 103 matching sequences were found. Following the molecular docking evaluation and considering the acting function and pathway of the plasminogen Kringle 5 in the human body, the most promising candidate was determined to be voltage-dependent anion channel-1 (VDAC-1), which was able to bind to Kringle 5 at -822.65 J·mol-1 of the binding energy at the residues of Lys12, Thr19, Ser57, Thr188, Arg139, Asn214, Ser240 and Lys274. A strong dose-dependent interaction occurred between the VDAC-1 and Kringle 5 (binding constant 2.43 × 103 L·mol-1) in SPR observation. Therefore, this study proposed that VDAC-1 was a potential ligand of plasminogen Kringle 5, and also demonstrated that the screening and validation of protein ligand using phage display peptide library with the molecular docking, along with SPR, was a practicable application.
authors.affiliation: College of Life Sciences, Northwest University, Xi'an 710069, P. R. China.|College of Biological Science and Engineering, Shaanxi University of Technology, Hanzhong 723000, P. R. China.|Shaanxi Province Key Laboratory of Bio-Resource, Shaanxi University of Technology, Hanzhong 723000, P. R. China.|Qinba Mountains of Bio-Resource Collaborative Innovation Center of Southern Shaanxi province, Shaanxi University of Technology, Hanzhong 723000, P. R. China.
authors.forename: Yin-Ku
authors.initials: YK
authors.lastname: Liang
authors.affiliation: College of Life Sciences, Northwest University, Xi'an 710069, P. R. China.
authors.forename: Liu-Jiao
authors.initials: LJ
authors.lastname: Bian
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D008024:Ligands; D062105:Molecular Docking Simulation; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D019151:Peptide Library; D010958:Plasminogen / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D016415:Sequence Alignment; D020349:Surface Plasmon Resonance; D050995:Voltage-Dependent Anion Channel 1 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D008024:Ligands; D010446:Peptide Fragments; D019151:Peptide Library; D011994:Recombinant Proteins; C494762:VDAC1 protein, human; C491499:plasminogen kringle 5; D010958:Plasminogen; D050995:Voltage-Dependent Anion Channel 1