title: Pharmacoproteomic analysis of a novel cell-permeable peptide inhibitor of tumor-induced angiogenesis.
abstract: P11, a novel peptide ligand containing a PDZ-binding motif (Ser-Asp-Val) with high affinity to integrin α(v)β(3) was identified from a hexapeptide library (PS-SPCL) using a protein microarray chip-based screening system. Here, we investigated the inhibitory mechanism of P11 (HSDVHK) on tumor-induced angiogenesis via a pharmacoproteomic approach. P11 was rapidly internalized by, human umbilical vein endothelial cells (HUVECs) via an integrin α(v)β(3)-mediated event. Caveolin and clathrin appeared to be involved in the P11 uptake process. The cell-penetrating P11 resulted in suppression of bFGF-induced HUVEC proliferation in a dose-dependent manner. Phosphorylation of extracellular-signal regulated kinase (ERK1/2) and mitogen-activated protein kinase kinase (MEK) in bFGF-stimulated HUVECs was inhibited by cell-permeable P11. Proteomic analysis via antibody microarray showed up-regulation of p53 in P11-treated HUVECs, resulting in induction of apoptosis via activation of caspases-3, -8, and -9. Several lines of experimental evidence strongly suggest that the molecular mechanism of P11, a novel anti-angiogenic agent, inhibits bFGF-induced HUVEC proliferation via mitogen-activated protein kinase kinase and extracellular-signal regulated kinase inhibition as well as p53-mediated apoptosis related with activation of caspases.
authors.affiliation: InnoPharmaScreen Inc, Hoseo University, Asan 336-795, Korea.
authors.forename: Ji-Young
authors.initials: JY
authors.lastname: Bang
authors.forename: Eung-Yoon
authors.initials: EY
authors.lastname: Kim
authors.forename: Dong-Ku
authors.initials: DK
authors.lastname: Kang
authors.forename: Soo-Ik
authors.initials: SI
authors.lastname: Chang
authors.forename: Moon-Hi
authors.initials: MH
authors.lastname: Han
authors.forename: Kwang-Hyun
authors.initials: KH
authors.lastname: Baek
authors.forename: In-Cheol
authors.initials: IC
authors.lastname: Kang
mesh_terms: D000595:Amino Acid Sequence; D017209:Apoptosis / Q000187:drug effects; D020169:Caspases / Q000378:metabolism; D049109:Cell Proliferation / Q000187:drug effects*; D057846:Cell-Penetrating Peptides / Q000378:metabolism / Q000494:pharmacology*; D002478:Cells, Cultured; D004789:Enzyme Activation; D020869:Gene Expression Profiling; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism; D006801:Humans; D039302:Integrin alphaVbeta3 / Q000037:antagonists & inhibitors / Q000378:metabolism; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism; D020935:MAP Kinase Signaling System / Q000187:drug effects; D009389:Neovascularization, Pathologic; D040081:Protein Array Analysis; D020543:Proteome / Q000378:metabolism; D040901:Proteomics; D014157:Transcription Factors / Q000378:metabolism
chemical_list: D057846:Cell-Penetrating Peptides; D039302:Integrin alphaVbeta3; D047908:Intracellular Signaling Peptides and Proteins; D020543:Proteome; D014157:Transcription Factors; C572394:p11 peptide; D020169:Caspases