RRC ID 63569
Author Hashimoto D, Semba S, Tsuda M, Kurokawa T, Kitamura N, Yasuda K, Gong JP, Tanaka S.
Title Integrin α4 mediates ATDC5 cell adhesion to negatively charged synthetic polymer hydrogel leading to chondrogenic differentiation.
Journal Biochem Biophys Res Commun
Abstract Negatively charged synthetic hydrogels have been known to facilitate various cellular responses including cell adhesion, proliferation, and differentiation; however, the molecular mechanism of hydrogel-dependent control of cell behavior remains unclear. Recently, we reported that negatively charged poly(2-acrylamido-2-methylpropanesulfonic acid) (PAMPS) gel induces chondrogenic differentiation of ATDC5 cells via novel protein reservoir function. In this study, we identified the cell adhesion molecules binding to PAMPS gels that act as mechanoreceptors. First, we performed a pull-down assay by particle gels using cell membrane proteins of ATDC5, and found that multiple membrane proteins bound to the PAMPS gel, whereas the uncharged poly(N,N'-dimethylacrylamide) gel as control did not bind to any membrane proteins. Western blot analysis indicated differential binding of integrin (ITG) isoforms to the PAMPS gel, in which the α4 isoform, but not α5 and αv, efficiently bound to the PAMPS gel. ITG α4 knockdown decreased cell spreading of ATDC5 on PAMPS gels, whereas the enhanced expression increased the behavior. Furthermore, ITG α4 depletion suppressed PAMPS gel-induced expression of bone morphogenic protein (BMP) 4 contributing to chondrogenic differentiation, in concordance with the reduction of ERK activation. These results demonstrated that membrane protein binding to PAMPS gels occurred in a charge-dependent manner, and that ITG α4 plays a crucial role in cell spreading on PAMPS gels and acts as a mechanoreceptor triggering cellular signaling leads to chondrogenic differentiation.
Volume 528(1)
Pages 120-126
Published 2020-7-12
DOI 10.1016/j.bbrc.2020.05.071
PII S0006-291X(20)30979-7
PMID 32456794
MeSH Animals Bone Morphogenetic Protein 4 / pharmacology Cell Adhesion / drug effects Cell Differentiation* / drug effects Cell Line Cell Movement / drug effects Chondrogenesis* / drug effects Extracellular Signal-Regulated MAP Kinases / metabolism Gene Knockdown Techniques Humans Hydrogels / chemistry* Integrin alpha4 / metabolism* Membrane Proteins / metabolism Mice Phosphorylation / drug effects Polymers / chemistry* Protein Binding / drug effects Sulfonic Acids / chemistry
IF 2.985
Human and Animal Cells ATDC5(RCB0565)