title: Small molecule inhibitors target the tissue transglutaminase and fibronectin interaction.
abstract: Tissue transglutaminase (TG2) mediates protein crosslinking through generation of ε-(γ-glutamyl) lysine isopeptide bonds and promotes cell adhesion through interaction with fibronectin (FN) and integrins. Cell adhesion to the peritoneal matrix regulated by TG2 facilitates ovarian cancer dissemination. Therefore, disruption of the TG2-FN complex by small molecules may inhibit cell adhesion and metastasis. A novel high throughput screening (HTS) assay based on AlphaLISA™ technology was developed to measure the formation of a complex between His-TG2 and the biotinylated FN fragment that binds TG2 and to discover small molecules that inhibit this protein-protein interaction. Several hits were identified from 10,000 compounds screened. The top candidates selected based on >70% inhibition of the TG2/FN complex formation were confirmed by using ELISA and bioassays measuring cell adhesion, migration, invasion, and proliferation. In conclusion, the AlphaLISA bead format assay measuring the TG2-FN interaction is robust and suitable for HTS of small molecules. One compound identified from the screen (TG53) potently inhibited ovarian cancer cell adhesion to FN, cell migration, and invasion and could be further developed as a potential inhibitor for ovarian cancer dissemination.
authors.affiliation: Indiana University Department of Medicine, Indiana University School of Medicine, Indianapolis, Indiana, United States of America.
authors.forename: Bakhtiyor
authors.initials: B
authors.lastname: Yakubov
authors.affiliation: Chemical Genomics Core Facility, Indiana University School of Medicine, Indianapolis, Indiana, United States of America.
authors.forename: Lan
authors.initials: L
authors.lastname: Chen
authors.affiliation: University of Maryland Department of Biochemistry and Molecular Biology, Indiana University School of Medicine, Indianapolis, Indiana, United States of America.
authors.forename: Alexey M
authors.initials: AM
authors.lastname: Belkin
authors.affiliation: Chemical Genomics Core Facility, Indiana University School of Medicine, Indianapolis, Indiana, United States of America.
authors.forename: Sheng
authors.initials: S
authors.lastname: Zhang
authors.affiliation: Indiana University Department of Medicine, Indiana University School of Medicine, Indianapolis, Indiana, United States of America.
authors.forename: Bhadrani
authors.initials: B
authors.lastname: Chelladurai
authors.affiliation: Indiana University Simon Cancer Center, Indiana University School of Medicine, Indianapolis, Indiana, United States of America ; Department of Biochemistry and Molecular Biology, Indiana University School of Medicine, Indianapolis, Indiana, United States of America.
authors.forename: Zhong-Yin
authors.initials: ZY
authors.lastname: Zhang
authors.affiliation: Indiana University Department of Medicine, Indiana University School of Medicine, Indianapolis, Indiana, United States of America ; Indiana University Simon Cancer Center, Indiana University School of Medicine, Indianapolis, Indiana, United States of America ; Department of Biochemistry and Molecular Biology, Indiana University School of Medicine, Indianapolis, Indiana, United States of America ; VA Roudebush Hospital, Indiana University School of Medicine, Indianapolis, Indiana, United States of America.
authors.forename: Daniela
authors.initials: D
authors.lastname: Matei
mesh_terms: D015153:Blotting, Western; D002448:Cell Adhesion / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005353:Fibronectins / Q000037:antagonists & inhibitors* / Q000378:metabolism; D019204:GTP-Binding Proteins / Q000037:antagonists & inhibitors* / Q000378:metabolism; D057166:High-Throughput Screening Assays; D006801:Humans; D010051:Ovarian Neoplasms / Q000188:drug therapy* / Q000378:metabolism* / Q000473:pathology; D000091345:Protein Glutamine gamma Glutamyltransferase 2; D060066:Protein Interaction Maps / Q000187:drug effects*; D054852:Small Molecule Libraries / Q000494:pharmacology*; D011503:Transglutaminases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D014407:Tumor Cells, Cultured; D014945:Wound Healing / Q000187:drug effects
chemical_list: D005353:Fibronectins; D054852:Small Molecule Libraries; D000091345:Protein Glutamine gamma Glutamyltransferase 2; D011503:Transglutaminases; D019204:GTP-Binding Proteins