RRC ID 54063
Author Png E, Hou A, Tong L.
Title Mechanistic role of transglutaminase-2 in focal adhesions.
Journal Sci Rep
Abstract Transglutaminase (TG)-2 interacts with matrix proteins and integrins, forming focal adhesions (FA) to initiate cell migration, thus playing a vital role in wound healing. Previously we showed that TG-2 influenced phosphorylation of paxillin and other FA proteins. Here, we aimed to investigate the molecular mechanism of TG-2 regulation of paxillin. Human corneal epithelial cells expressing shRNA against TG-2 (shTG) and scrambled sequence control (shRNA) were cultured. TG-2 was pulled down by anti-paxillin antibody, but not MAP3K12. Cell-free interaction assay with immobilized paxillin shows that TG-2 bind to paxillin directly. JNK was the strongest kinase for paxillin phosphorylation in the in-vitro kinase screen, but TG-2 could not phosphorylate paxillin directly. Increasing TG-2 concentrations did not increase the amount of JNK in the TG-2/paxillin complex. Immunofluoresent staining shows that TG-2 colocalises with vinculin and paxillin in FA of migrating cells. TG-2 binds to paxillin and JNK-containing FA but does not recruit JNK directly. Taken together with previous findings, TG-2 binds paxillin non-covalently, and JNK can phosphorylate paxillin, these processes critically regulate corneal epithelial adhesion and migration.
Volume 8(1)
Pages 12370
Published 2018-8-17
DOI 10.1038/s41598-018-30172-8
PII 10.1038/s41598-018-30172-8
PMID 30120307
PMC PMC6098073
MeSH Blotting, Western Cell Line Cell Movement / genetics Cell Movement / physiology Focal Adhesions / genetics Focal Adhesions / metabolism* GTP-Binding Proteins / genetics GTP-Binding Proteins / metabolism* Humans Immunoprecipitation MAP Kinase Signaling System / genetics MAP Kinase Signaling System / physiology Mass Spectrometry Paxillin / genetics Paxillin / metabolism Phosphorylation / genetics Phosphorylation / physiology Protein Binding Protein Glutamine gamma Glutamyltransferase 2 RNA, Small Interfering Transglutaminases / genetics Transglutaminases / metabolism*
IF 3.998
Times Cited 1
Human and Animal Cells HCE-T(RCB2280)