Modified BIGH3 with an RGDRGD motif promotes human corneal epithelial cell adhesion and migration in vitro.
OBJECTIVE
BIGH3 protein plays an important role in mediating human corneal epithelial (HCE) cell adhesion and migration. The aim of this study was to investigate the effects of native and modified BIGH3 protein containing an Arg-Gly-Asp-Arg-Gly-Asp (RGDRGD) motif on the adhesion and migration of HCE cells.


METHODS
A modified human BIGH3 gene containing an RGDRGD motif was obtained by rapid site-directed mutagenesis. Recombinant human native and modified BIGH3 proteins were then obtained and purified. The effects of the native and the modified version on the adhesion and migration of HCE cells were examined in the presence or absence of anti-alpha3beta1 antibody or anti-BIGH3 antibody or RGD peptide in vitro.


RESULTS
Recombinant native and modified BIGH3 proteins were successfully obtained and significantly promoted the adhesion and migration of human HCE cells in vitro, and the construct with the RGDRGD motif was more effective. The enhanced adhesion and migration were blocked by anti-alpha3beta1antibody or anti-BIGH3 antibody or RGD peptide.


CONCLUSIONS
BIGH3 promotes HCE cell adhesion and migration; modified RGDRGD-BIGH3 was more effective than native BIGH, and this is mediated by the Arg-Gly-Asp (RGD) motif and alpha3beta1integrin in a RGD-dependent manner.
Ge
Hongyan
H
Eye Hospital, The First Clinical Hospital, Harbin Medical University, Harbin, China.
Cao
Wenping
W
Leng
Fei
F
Chen
Weiye
W
Liu
Ping
P
Chen
Weiye
W
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D002448:Cell Adhesion / Q000502:physiology*; D018929:Cell Culture Techniques; D002465:Cell Movement / Q000502:physiology*; D019573:Epithelium, Corneal / Q000502:physiology*; D016326:Extracellular Matrix Proteins / Q000302:isolation & purification / Q000502:physiology*; D015870:Gene Expression; D006801:Humans; D039201:Integrin alpha3beta1; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009842:Oligopeptides / Q000737:chemistry / Q000502:physiology*; D010957:Plasmids; D017510:Protein Folding; D011994:Recombinant Proteins / Q000302:isolation & purification; D016212:Transforming Growth Factor beta / Q000302:isolation & purification / Q000502:physiology*; D000099222:betaIG-H3 Protein
D016326:Extracellular Matrix Proteins; D039201:Integrin alpha3beta1; D009842:Oligopeptides; D011994:Recombinant Proteins; D016212:Transforming Growth Factor beta; D000099222:betaIG-H3 Protein; C047981:arginyl-glycyl-aspartic acid